Compositions and methods for the treatment of disorders related to glucosylceramidase beta 1 deficiency

Abstract

The disclosure relates to compositions and methods for, inter alia, altering, e.g., enhancing, the level of GBA1 protein via delivery using an adeno-associated viral (AAV) capsid variant. The compositions and methods of the present disclosure are useful, inter alia, for the treatment of subjects who have, have been diagnosed with having, or are at risk of having a GBA1-related disorder, e.g., Parkinson's Disease (PD), Gaucher Disease (GD), Parkinson's Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), or Lewy Body Dementia (LBD).

Description

COMPOSITIONS AND METHODS FOR THE TREATMENT OF DISORDERSRELATED TO GLUCOSYLCERAMIDASE BETA 1 DEFICIENCYRELATED APPLICATIONS

[0001] This application claims the benefit of and priority to US Provisional Application Serial No. 63 / 606,162, filed December 5, 2023, the contents of which are incorporated herein by reference in their entirety.SEQUENCE LISTING

[0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing file, entitled 14640_0101 -00304_SL.xml. was created on November 18, 2024, and is 1,936.273 bytes in size. The information in electronic format of the Sequence Listing is incorporated herein by reference in its entirety7.FIELD

[0003] Described herein are compositions and methods relating to adeno-associated virus (AAV) viral particles for the delivery of polynucleotides, e.g.. polynucleotides encoding glucosylceramidase beta 1 (GBA1) proteins and peptides for use in the treatment of GBAl-related disorders, which include Parkinson’s Disease (PD) and other GBAl-related disorders, including Gaucher Disease, Parkinson’s Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), and Lewy Body Dementia (LBD). In some embodiments, compositions described herein may be used to treat a subject in need thereof, such as a human subject diagnosed with a GBAl-related disorder or other condition resulting from a deficiency in the quantity and / or function of GBA1 protein.BACKGROUND

[0004] Lysosomal acid glucosylceramidase, commonly called glucosylcerebrosidase or Gcase, a D- glucosyl-N-acylsphingosine glucohydrolase, is a lysosomal membrane protein important in glycolipid metabolism. The enzyme is encoded by the glucosylceramidase beta 1 (GBA1) gene (Ensembl Gene ID No. ENSG00000177628). This enzyme, together with Saposin A and Saposin C, catalyzes the hydrolysis of glucosylceramide to ceramide and glucose. See Vaccaro, Anna Maria, et al. Journal of Biological Chemistry m.Tl (1997): 16862-16867.

[0005] Mutations in GBA are known to cause disease in human subjects. Homozygous or compound heterozygous GBA1 mutations lead to Gaucher disease ("GD”). See Sardi, S. Pablo, Jesse M.Cedarbaum. and Patrik Brundin. Movement Disorders 33.5 (2018): 684-696. Gaucher disease is one of the most prevalent lysosomal storage disorders, with an estimated standardized birth incidence in the general population of 0.4 to 5.8 individuals per 100,000. Heterozygous GBA1 mutations can lead to PD. Indeed, GBA1 mutations occur in 7-10% of total PD patients, making GBA1 mutations the most important genetic risk factor of PD. PD-GBA patients have reduced levels of the lysosomal enzyme beta-glucocerebrosidase (Gcase), which results in increased accumulations of glycosphingolipid glucosylceramide (GluCer), which in turn is correlated with exacerbated a-Synuclein aggregation and concomitant neurological symptoms. GD and PD, as well as other lysosomal storage disorders or Lewy body diseases such as Lewy Body Dementia (LBD). See Sidransky, E. and Lopez, G. Lancet Neurol. 2012 November; 11(11): 986-998.

[0006] To date, there are limited available treatments for GBAl-related disorders such as PD, and delivery to the adult central nervous system (CNS) remains a significant challenge in the development of new and effective therapies. Thus, there remains a long-felt need to develop pharmaceutical compositions and methods that can be delivered to the CNS for the treatment of PD and other GBAl-related disorders and to ameliorate deficiencies of GBA1 protein in subjects, e.g., human subjects, afflicted with GBAl- related disorders.

[0007] Adeno-associated viruses (AAVs) have emerged as a widely studied and utilized viral particles for delivery of therapeutically effective polypeptides to mammalian cells. See, e.g.. Tratschin et al.. Mol. Cell Biol., 5(11):3251-3260 (1985) and Grimm et al.. Hum. Gene Then, 10(15):2445-2450 (1999).

[0008] The present disclosure provides improved pharmaceutical compositions and methods. In some embodiments, the disclosure provides methods of treatment using AAV capsid variants that are capable of delivering GBA1 to a target cell or tissue, e.g., a CNS cell or tissue.SUMMARY

[0009] In some aspects, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV capsid variant and a viral genome, wherein the viral genome comprises a glucosylceramidase beta 1 (GBAl)-encoding sequence and the AAV capsid variant comprises an amino acid sequence having the formula [N 1 ]-[N2]-[N3] (SEQ ID NO: 4681) in loop VIII. wherein [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682); [N 1] comprises amino acids Xi, X2, X3. and X4, wherein X4is Q. K, E, S. P. R, N. or H; and / or [N3] comprises amino acids Xs, X6, and X7. wherein Xs is I, V, T, M, S, N, L, or F. In some embodiments, the AAV capsid variant is an AAV9 capsid variant. In some embodiments, the AAV capsid variant comprises an amino acid sequence that is at least 95% identical to positions 203-736 of the amino acid sequence of SEQ ID NO: 981. In some embodiments, loop VIII is present at amino acids comprising those corresponding to positions 580-599 of the amino acid sequence of SEQ ID NO: 981.

[0010] In some embodiments. X] is T. S, R, A, I. C, N. K, L, or Q; X2is N, T, G, V, S. Y, K, I, H, D, or F; and X3is T, N, K. D, I, S, P. A, Y, E, V, L. M, R. H, Q, or C. In some embodiments, X6is A, Y. P, N, S, T, G, E, V, W, F, or Q; and X7is Q, G, N, K. H, R, E, L, P, or M.[OH] In some embodiments, [Nl] comprises the amino acid sequence of TNTQ (SEQ ID NO: 4688). In some embodiments, [N3] comprises the amino acid sequence of IAQ. In some embodiments, [N1]-N2]-[N3] comprises the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343).

[0012] In some embodiments, [Nl] is present at amino acids corresponding to positions 582-585 of the amino acid sequence of SEQ ID NO: 981, [N2] is present at amino acids corresponding to positions 586-589 of the amino acid sequence of SEQ ID NO: 981, and [N3] is present at amino acids corresponding to positions 590-592 of the amino acid sequence of 981.

[0013] hi some embodiments, the AAV capsid variant further comprises [N4] comprising amino acids X8, X9, Xio, and Xu, wherein: X8is T. S, N, P, A, or I; X9is G, N, D, R. V, A, S, or Q; Xiois W, S, C, R, L. or G; and / or Xu is V, A, S, I, C, G. D, F, L. or T. In some embodiments, [N4] comprises the amino acid sequence of TGWV (SEQ ID NO: 5066). In some embodiments, [N4] is present at amino acids corresponding to positions 593-596 of the amino acid sequence of ID NO: 981.

[0014] In some embodiments, [N1]-[N2]-[N3]-[N4] comprises the amino acid sequence of TNTQDWHRIAQTGWV (SEQ ID NO: 201).

[0015] In some aspects, the present disclosure provides an AAV particle comprising a viral genome comprising a GBA1 -encoding sequence and an AAV capsid variant comprising an amino acid sequence that is at least 95% identical to positions 203-736 of the amino acid sequence of SEQ ID NO: 981, wherein the AAV capsid variant comprises: T at an amino acid corresponding to position 584 of the amino acid sequence of SEQ ID NO: 981, D at an amino acid corresponding to position 586 of the amino acid sequence of SEQ ID NO: 981, W at an amino acid corresponding to position 587 of the amino acid sequence of SEQ ID NO: 981, H at an amino acid corresponding to position 588 of the amino acid sequence of SEQ ID NO: 981, R at an amino acid corresponding to position 589 of the amino acid sequence of SEQ ID NO: 981, and I at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 981. In some embodiments, the AAV capsid variant comprises an amino acid sequence that is at least 95% identical to positions 138-736 of the amino acid sequence of SEQ ID NO: 981.

[0016] In some aspects, the present disclosure provides an AAV particle comprising a viral genome comprising a GBA1 -encoding sequence and an AAV capsid variant comprising an amino acid sequence that is at least 95% identical to positions 138-736 of the amino acid sequence of SEQ ID NO: 981, wherein the AAV capsid variant comprises: T at an amino acid corresponding to position 584 of the amino acid sequence of SEQ ID NO: 981. D at an amino acid corresponding to position 586 of the amino acid sequence of SEQ ID NO: 981, W at an amino acid corresponding to position 587 of the amino acid sequence of SEQ ID NO: 981, H at an amino acid corresponding to position 588 of the amino acid sequence of SEQ ID NO: 981, R at an amino acid corresponding to position 589 of the amino acid sequence of SEQ ID NO: 981, and I at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 981. In some embodiments, the AAV capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 981.

[0017] In some aspects, the present disclosure provides an AAV particle comprising a viral genome comprising a GBA1 -encoding sequence and an AAV capsid variant comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 981, wherein the AAV capsidvariant comprises: T at an amino acid corresponding to position 584 of the amino acid sequence of SEQ ID NO: 981, D at an amino acid corresponding to position 586 of the amino acid sequence of SEQ ID NO: 981, W at an amino acid corresponding to position 587 of die amino acid sequence of SEQ ID NO: 981, H at an amino acid corresponding to position 588 of the amino acid sequence of SEQ ID NO: 981, R at an amino acid corresponding to position 589 of the amino acid sequence of SEQ ID NO: 981, and I at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 981.

[0018] In some aspects, the present disclosure provides an AAV particle comprising a viral genome comprising a GBA1 -encoding sequence and an AAV capsid variant, wherein the AAV capsid variant comprises: (i) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 981; (ii) an amino acid sequence that is at least 99% identical to positions 138-736 of the amino acid sequence of SEQ ID NO: 981; and / or (iii) an amino acid sequence that is at least 99% identical to positions 203-736 of the amino acid sequence of SEQ ID NO: 981. In some embodiments, the AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941).

[0019] In some embodiments, the AAV capsid variant comprises at least 8, at least 9, or at least 10 consecutive amino acids from the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343). In some embodiments, the AAV capsid variant comprises the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343) present at amino acids corresponding to positions 582-592 of the amino acid sequence of SEQ ID NO: 981. In some embodiments, the amino acid sequence of TQDWHRI (SEQ ID NO: 941) or TNTQDWHRIAQ (SEQ ID NO: 343) is present in loop VIII, wherein loop VIII comprises amino acids 580-599 of the amino acid sequence of SEQ ID NO: 981.

[0020] In some embodiments, the AAV capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 981; (ii) the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981; and / or (iii) the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

[0021] In some embodiments, the viral genome of the AAV particle encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1775, 1740, 1742, 1744, 1746, or 1748 or an amino acid sequence that is at least 90% (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) identical thereto. In some embodiments, the viral genome of the AAV particle encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1775 or an amino acid sequence that is at least 90% identical (e g., at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto. In some embodiments, the viral genome encodes a wildtype GBA1 protein. In some embodiments, the viral genome encodes a human GBA1 protein, a dog GBA1 protein, an equine GBA1 protein, or a monkey GBA1 protein. In some embodiments, the viral genome encodes a wildtype human GBA1 protein. In some embodiments, the viral genome does not encode a hemagglutinin (HA) tag. In some embodiments, the GBA1 -encoding sequence comprises SEQ ID NO: 2002 or SEQ ID NO: 1773, or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. atleast 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto. In some embodiments, the viral genome further comprises a signal sequence-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2005 or SEQ ID NO: 1850, or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto. In some embodiments, the viral genome comprises the nucleotide sequence of 2001 or SEQ ID NO: 1772, or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto. In some embodiments, the viral genome encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1774 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto.

[0022] In some embodiments, the viral genome comprises a promoter operably linked to the GBA1- encoding sequence. In some embodiments, the promoter comprises a human elongation factor 1 alpha (EF-la) promoter, a chicken f> -actin (CBA) promoter, a CAG promoter, a CAG derivative promoter, a CMV immediate-early enhancer and / or promoter, a CMV promoter, a glucuronidase (GUSB) promoter, a ubiquitin C (UBC) promoter, a neuron-specific enolase (NSE) promoter, a platelet-derived growth factor (PDGF) promoter, a platelet-derived growth factor B-chain (PDGF-0) promoter, an intercellular adhesion molecule 2 (ICAM-2) promoter, a synapsin (Syn) promoter, a methyl-CpG binding protein 2 (MeCP2) promoter, a Ca2+ / calmodulin-dependent protein kinase II (CaMKII) promoter, a metabotropic glutamate receptor 2 (mGluR2) promoter, a neurofilament light chain (NFL) promoter, a neurofilament heavy chain (NFH) promoter, a 0-globin minigene n02 promoter, a preproenkephalin (PPE) promoter, an enkephalin (Enk) and excitatory' amino acid transporter 2 (EAAT2) promoter, a glial fibrillary' acidic protein (GFAP) promoter, a myelin basic protein (MBP) promoter, a cardiovascular promoter (e.g., aMHC, cTnT, and CMV-MLC2k), a liver promoter (e.g., hAAT, TBG), a skeletal muscle promoter (e.g., desmin, MCK, C512), or a fragment, e.g., a truncation, or a functional variant thereof. In some embodiments, the promoter comprises a CBA promoter. In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

[0023] In some embodiments, the viral genome further comprises an enhancer. In some embodiments, the enhancer comprises a CMV immediate-early (CMVie) enhancer. In some embodiments, the enhancer comprises the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

[0024] In some embodiments, the viral genome further comprises an intron. In some embodiments, the intron comprises the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is atleast 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

[0025] In some embodiments, the viral genome further comprises a polyadenylation (poly A) region. In some embodiments, the polyA region comprises the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

[0026] In some embodiments, the viral genome further comprises an inverted terminal repeat (ITR). In some embodiments, the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto. In some embodiments, the viral genome comprises a 5’ ITR and a 3’ ITR. wherein the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto, and the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%. at least 97%, at least 98%. or at least 99% identical) thereto.

[0027] In some embodiments, the viral genome further comprises a nucleotide sequence encoding one or more miR183 binding sites. In some embodiments, the viral genome encodes at least four miR183 binding sites. In some embodiments, each of the at least four miR183 binding sites is separated by a spacer. In some embodiments, each of the miR183 binding sites is encoded by a nucleotide sequence that comprises the nucleotide sequence of SEQ ID NO: 1847 or a nucleotide sequence that has up to three modifications relative thereto.

[0028] In some embodiments, the viral genome further comprises a nucleotide sequence encoding a miR183 binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

[0029] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) a promoter; (iii) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; and (iv) a 3 ’ ITR.

[0030] In some embodiments, the viral genome comprises (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer; (iii) a promoter: (iv) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequencethat is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and (v) a 3‘ ITR.

[0031] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer; (iii) a promoter: (iv) an intron; (v) the GBAl-encoding sequence, wherein the GBA1- encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and (vi) a 3 ’ ITR.

[0032] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; and (vi) a 3’ ITR.

[0033] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; and (vii) a 3’ ITR.

[0034] In some embodiments, the viral genome comprises: (i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%. at least 97%. at least 98%, or at least 99% identical) thereto;(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBA1- encoding sequences comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; and (viii) a 3 ’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

[0035] In some embodiments, (i) the 5' ITR comprises the nucleotide sequence of SEQ ID NO: 1829; (ii) the enhancer comprises the nucleotide sequence of SEQ ID NO: 1831; (iii) the promoter comprises the nucleotide sequence of SEQ ID NO: 1834; (iv) the intron comprises tire nucleotide sequence of SEQ ID NO: 1842; (v) the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001; (vi) the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846; and (viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

[0036] In some embodiments, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto. In some embodiments, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006. In some embodiments, the viral genome consists of the nucleotide sequence of SEQ ID NO: 2006.

[0037] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) a promoter; (iii) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises tire nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) a nucleotide sequence encoding at least one miR183 binding site; and (v) a 3’ ITR.

[0038] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) a promoter; (iii) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; (iv) a nucleotide sequenceencoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and (v) a 3’ ITR.

[0039] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer; (iii) a promoter: (iv) the GBAl-encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and (vi) a 3’ ITR.

[0040] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer; (iii) a promoter: (iv) an intron; (v) the GBAl-encoding sequence, wherein the GBAl- encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and (vii) a 3’ ITR.

[0041] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and (vii) a 3‘ ITR.

[0042] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR); (ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBAl-encodingsequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) a nucleotide sequence encoding a miR183 binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and (vii) a 3’ ITR.

[0043] In some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; (vi) a nucleotide sequence encoding at least one miR183 binding site, wherein the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of SEQ ID NO: 1847; (vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and (viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

[0044] hi some embodiments, the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; (iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%. at least 97%, at least98%, or at least 99% identical) thereto; (v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) a nucleotide sequence encoding a miR183 binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; and (viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

[0045] In some embodiments, (i) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829; (ii) the enhancer comprises the nucleotide sequence of SEQ ID NO: 1831; (iii) the promoter comprises the nucleotide sequence of SEQ ID NO: 1834; (iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842; (v) the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001; (vi) the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849; (vii) the polyA region comprises the nucleotide sequence of SEQ ID NO: 1846; and (viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

[0046] In some embodiments, the viral genome comprises viral genome comprises the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto. In some embodiments, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007. In some embodiments, the viral genome consists of the nucleotide sequence of SEQ ID NO: 2007.

[0047] In some aspects, the present disclosure provides a cell comprising an AAV particle provided herein. In some embodiments, the cell is a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

[0048] In some aspects, the present disclosure provides a method of making an AAV particle provided herein, the method comprising: (i) providing a cell comprising viral genome comprising a GBAl-encoding sequence and a nucleic acid encoding an AAV capsid variant; and (ii) incubating the cell under conditions suitable to encapsulate the viral genome in the AAV capsid variant; thereby making the AAV particle.

[0049] In some embodiments of a method of making an AAV particle, the viral genome comprises (i) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%. at least 93%. at least 94%, at least 95%. at least 96%, atleast 97%. at least 98%, or at least 99% identical) thereto; or (ii) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at that is least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and the AAV capsid variant comprises (a) the amino acid sequence of SEQ ID NO: 981 or an amino acid sequence that is at least 90% identical (e g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (b) the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981 or an amino acid sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; and / or (c) the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%. at least 92%, at least 93%. at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto.

[0050] In some embodiments of a method of making an AAV particle, the viral genome comprises: (i) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; or (ii) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; and the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981, the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981, and / or the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981. In some embodiments of a method of making an AAV particle, the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or SEQ ID NO: 2007 and the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981, the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981, and / or the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

[0051] In some embodiments, the method of making an AAV particle comprises, prior to step (i), introducing a nucleic acid molecule comprising the viral genome into the cell. In some embodiments, the method of making an AAV particle comprises, prior to step (i). introducing the nucleic acid encoding the AAV capsid variant into the cell. In some embodiments, the cell comprises a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an S19 cell), or a bacterial cell.

[0052] In some aspects, the present disclosure provides a pharmaceutical composition comprising an AAV particle provided herein and a pharmaceutically acceptable excipient.

[0053] In some aspects, the present disclosure provides a method of delivering an AAV particle encoding a GBA1 protein to cell, comprising administering an effective amount of a pharmaceutical composition or AAV particle provided herein. In some embodiments, the cell is in a subject. In someembodiments, the subject, has, has been diagnosed with having, or is at risk of having a GBAl-related disorder.

[0054] In some aspects, the present disclosure provides a method of treating a GBAl-related disorder in a subject, comprising administering to the subject an effective amount of a pharmaceutical composition or AAV particle provided herein. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having the GBAl-related disorder.

[0055] In some embodiments, the GBAl-related disorder is a GBAl-related neurodegenerative or neuromuscular disorder. In some embodiments, the GBAl-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, GD type 2, or GD type 3), Dementia with Lewy Bodies (DLB). Lewy Body Dementia (LBD), Multiple System Atrophy (MSA). Alzheimer’s Disease (AD). Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome. In some embodiments, the GBAl-related disorder is PD. In some embodiments, the GBA-1 related disorder is LBD. In some embodiments, the GBAl-related disorder is DLB. In some embodiments, the GBAl- related disorder is GD (e.g.. GD type 1, GD type 2. or GD type 3).

[0056] In some aspects, the present disclosure provides a method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Parkinson’s Disease (PD), comprising administering to the subject an effective amount of a pharmaceutical composition or AAV particle provided herein. In some embodiments, the subject has. has been diagnosed with having, or is at risk of having PD.

[0057] In some aspects, the present disclosure provides a method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Lewy Body Dementia (LBD), comprising administering to the subject an effective amount of a pharmaceutical composition or AAV particle provided herein. In some embodiments, tire subject has, has been diagnosed with having, or is at risk of having LBD.

[0058] In some aspects, the present disclosure provides a method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Dementia with Lewy Bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition or AAV particle provided herein. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having DLB.

[0059] In some aspects, the present disclosure provides a method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Gaucher Disease (GD), comprising administering to the subject an effective amount of a pharmaceutical composition or AAV particle provided herein. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having GD (e.g.. GD type 1, GD type 2. or GD type 3). In some embodiments, the GD is GD type 1. In some embodiments, the GD is GD type 2. In some embodiments, the GD is GD type 3.

[0060] In some embodiments, the subject has one or more mutations in the GBA1 gene.

[0061] In some embodiments, the subject has lower GCase activity prior to administration of the pharmaceutical composition or AAV particle as compared to GCase activity in an individual who does not have a GBA1 -related disorder. In some embodiments, the level of GCase activity' is measured by a 4- MUG assay or a SensoLyte Blue Glucocerebrosidase assay.

[0062] hi some embodiments, the treating results in the prevention of progression of a GBA-related disorder in the subject. In some embodiments, the treating results in amelioration of at least one symptom of the GBA1 -related disorder in the subject. In some embodiments, the at least one symptom comprises developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, generalized weakness, trembling of a limb, depression, visual hallucinations, cognitive decline, or a combination thereof. In some embodiments, the treating results in a change in one or more biomarkers comprising a GCase activity, a level of glucocerebroside and other glycolipids, (e.g.. within immune cells such as macrophages), a level of synuclein aggregates (e.g., Lewy bodies), or a combination thereof.

[0063] In some embodiments, the subject is a human.

[0064] In some embodiments, the AAV particle or the pharmaceutical composition is delivered to a cell or tissue of the central nervous system (CNS) in the subject. In some embodiments, the cell or tissue of the CNS is a cell or tissue of the amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region).

[0065] In some embodiments, the AAV particle or the pharmaceutical composition is delivered to a peripheral cell or tissue in the subject. In some embodiments, the peripheral cell or tissue is a cell or tissue of the heart, skeletal muscle, sympathetic ganglia, and / or plasma.

[0066] In some embodiments, the AAV particle or pharmaceutical composition is delivered to the subject via intravenous administration.

[0067] hi some embodiments, a method of delivery or treating provided herein further comprises evaluating, e.g., measuring, the level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) in the subject, e.g., in a cell, tissue, or fluid of the subject. In some embodiments, the level of GBA1 protein expression is measured by an enzy me-linked immunosorbent assay (ELISA), a Western blot, or an immunohistochemistry assay. In some embodiments, evaluating the subject’s level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) is performed prior to and / or subsequent to administration of the pharmaceutical composition or AAV particle. In some embodiments, the subject’s level of GBA1 expression (e.g., GBA1 gene expression. GBA1 mRNA expression, and / or GBA1 proteinexpression) prior to administration is compared to the subject’s level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) subsequent to administration. In some embodiments, the method comprises evaluating the level of GBA1 expression in a cell or tissue of the central nervous system. In some embodiments, the subject’s level of GBA1 protein expression subsequent to administration is increased relative to the subject's level of GBA1 protein expression prior to administration. In some embodiments, the method further comprises evaluating, e.g., measuring, the level of GCase activity in the subject.

[0068] In some embodiments, administering to the subject a pharmacal composition or AAV particle provided herein results in an increase in: (i) GCase activity in a cell, tissue, (e.g., a cell or tissue of the CNS, e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)), and / or fluid (e.g.. CSF and / or serum) of the subject relative to baseline and / or relative to GCase activity in a cell, tissue, or fluid of an individual with a GBA1 -related disorder who has not been administered the pharmaceutical composition or AAV particle; (ii) the number and / or level of viral genomes (VG) per cell in a CNS tissue (e.g.. amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gy rus, hippocampus, inferior colliculus, inferior olivary7complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)) of the subject relative to the number and / or level of VG per cell in a peripheral tissue of the subject; and / or (iii) GBA1 mRNA expression in a cell or tissue (e.g., a cell or tissue of the CNS, e.g.. amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory7cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus. oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)) of the subject relative to baseline and / or relative to GBA1 mRNA expression in a cell or tissue of an individual with a GBAl-related disorder who has not been administered the pharmaceutical composition or AAV particle.

[0069] In some embodiments, a method of delivery or treatment provided herein further comprises administering to the subject at least one additional agent and / or therapy. In some embodiments, the atleast one additional agent and / or therapy comprises an agent and / or therapy suitable for treating a GBA1- related disorder. In some embodiments, the at least one additional agent and / or therapy comprises enzy me replacement therapy (ERT) (e.g., imiglucerase, velaghicerase alfa, or taliglucerase alfa); substrate reduction therapy (SRT) (e.g., eliglustat or miglustat), levodopa, carbidopa, Safinamide, a dopamine agonist (e.g., pramipexole, rotigotine, or ropinirole), a dopamine antagonist (e.g., quetiapine, clozapine), an anticholinergic (e.g., benztropine or trihexyphenidyl), a cholinesterase inhibitor (e.g., rivastigmine, donepezil, or galantamine), an N-methyl-d-aspartate (NMD A) receptor antagonist (e.g., memantine), or a combination thereof. In some embodiments, the at least one additional agent and / or therapy is suitable for treating Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1. GD type 2, or GD type 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD). Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1). or Hallervorden-Spatz Syndrome. In some embodiments, the at least one additional agent and / or therapy is suitable for treating PD, LBD, GD. or DLB. In some embodiments, the GD is GD type 1, GD type 2, or GD ty pe 3.

[0070] In some embodiments, a method of delivery or treatment provided herein further comprises administering an immunosuppressant to the subject. In some embodiments, the immunosuppressant comprises a corticosteroid (e.g.. prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxy chloroquine. In some embodiments, a method of delivery or treatment provided herein further comprises administering a blood transfusion to the subject.

[0071] In some aspects, the present disclosure provides a pharmaceutical composition or AAV particle provided herein for use in a method of treating a disorder provided herein.

[0072] In some aspects, the present disclosure provides a pharmaceutical composition or AAV particle provided herein for use in the treatment of a GBAl-rclatcd disorder in a subject. In some embodiments, the GBAl-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, GD type 2, or GD type 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome. In some embodiments, the GBAl- related disorder is PD. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having PD. In some embodiments, the GBAl-related disorder is LBD. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having LBD. In some embodiments, the GBAl-related disorder is DLB. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having DLB. In some embodiments, the GBAl-related disorder is GD (e.g.. GD ty pe 1, GD type 2, or GD type 3). In some embodiments, the subject has, has been diagnosed with having, or is at risk of having GD (e.g., GD type 1, GD type 2, or GD ty pe 3).

[0073] In some aspects, the present disclosure provides a use of a pharmaceutical composition or AAV particle provided herein in the manufacture of a medicament for the treatment of a GBA1 -related disorder in a subject. In some embodiments, the GBA1 -related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, GD type 2, or GD type 3), Dementia with Lewy Bodies (DLB). Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD). Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome. In some embodiments, the GBAl-related disorder is PD. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having PD. In some embodiments, the GBAl-related disorder is GD. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having GD. In some embodiments, the GD is GD type 1. In some embodiments, the GD is GD type 2. In some embodiments, the GD is GD type 3. In some embodiments, the GBAl-related disorder is LBD. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having LBD. In some embodiments, the GBAl-related disorder is DLB. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having DLB.Enumerated Embodiments1. An adeno-associated virus (AAV) particle comprising a viral genome comprising a P- glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant), wherein tire AAV capsid variant comprises an amino acid sequence having the following formula: [N1]-[N2]-[N3] (SEQ ID NO: 4681), wherein [N2] comprises die amino acid sequence of DWHR (SEQ ID NO: 4682) and wherein:(i) [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4 is Q, K, E, S, P, R, N, H; and / or(ii) [N3] comprises amino acids X5, Xg, and X-, wherein X, is I, V, T, M, S, N, L, F.2. The AAV particle of embodiment 1, wherein the AAV capsid variant comprises the amino acid Q at position 585, as numbered according to SEQ ID NO: 138 or 981.3. The AAV particle of embodiment 1, wherein the AAV capsid variant comprises an amino acid other than Q at position 585. as numbered according to SEQ ID NO: 138 or 981.4. The AAV particle of embodiment 1 or 3, wherein the AAV capsid variant comprises the amino acid K at position 585, as numbered according to SEQ ID NO: 138 or 981.5. The AAV particle of any one of embodiments 1-4. wherein the AAV capsid variant comprises one. two. three, or all of:(i) an amino acid other than T at position 582 (e.g.. S, R, A. I. C, N, K, L, or Q);(ii) an amino acid other than N at position 583 (e.g., T, G, V, S. Y, K, I, H, D, or F);(iii) an amino acid other than H at position 584 (e.g., T, N, K, D, I, S, P, A, Y, E, V, L. M, R, Q, or C); and / or(iv) an amino acid other than Q at position 585 (e.g., K, E, S, P, R, N, or H); wherein the amino acids are numbered according to SEQ ID NO: 138.6. The AAV particle of any one of embodiments 1-5, wherein [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4is Q, K, E, S, P, R, N. or H.7. The AAV particle of embodiment 6, wherein X4is Q or K.8. The AAV particle of any one of embodiment 7. wherein X4is Q.9. The AAV particle of embodiment 7, wherein X4is K.10. The AAV particle of any one of embodiments 1-9, wherein the AAV capsid variant comprises an amino acid other than H at position 584 (e.g., T). as numbered according to SEQ ID NO: 138.11. The AAV particle of any one of embodiments 1-10, wherein the AAV capsid variant comprises the amino acid T at position 584, as numbered according to SEQ ID NO: 138 or 981.12. The AAV particle of any one of embodiments 1-11, wherein:(i) Xj is T, S, R, A. I. C, N, K, L, or Q;(ii) X2is N, T, G, V, S, Y, K, I, H, D, or F; and / or(iii) X3is T, N, K, D, I, S, P, A, Y, E, V, L, M, R, H, Q, or C.13. The AAV particle of any one of embodiments 1-12, wherein [Nl] comprises TN, NT, NK, SN, TT, RN, TG, TV, ST, TS, TY, AN, TK, TI, IN, TH, TD, CN, NN, KN, LN, SG, TF, RT, SY, SS, QN, ND, NP, GK, TA, VK, NY. TE, SK, Nl. YN, GT, TL, TM, YT. TR, NS, IT, NA, KT, GN, HT, DT. NE, NH, YI, HN. NQ, FS, NM, NL, SM, NC, VT, KQ, TQ. DQ, IQ, SQ, PS, KE, AQ, YQ, TP, EQ, VQ, LQ, MQ, KS, IE, RQ, IK, AK. PK, NR, HQ, QQ. or CQ.14. The AAV particle of any one of embodiments 1-13, wherein [Nl] comprises TNT, TNK, TNN, SNN, SNK, SNT, TTN, TND, TTL RNT. TTK, TTS, TTD, TNP, TTT. TGK. TTA. TVK. TNY. STK, TTE, TSK, TNI, TYN, STI, TTV, TGT, TTL, TTM, ANN, SNI. TKN. TYT. TTR, TNS, TST, TIT, INT, TNA, TKT, STN, ANT, RNN, TGN, TSN, THT, TDT, TNE, CNT, INN. NNN. KNN, LNN, TIN. TNH. STT, SNS, STS, TYI. SGT, THN, TNQ, RNI, TFS. RNS. TNM. RTT. KNT. TNL. TSM. SYT, TNC, SST, TVT. QNT. NTK. NNQ. NKQ, NNE, NTQ, NDQ, TIQ. TKQ. TSQ. TDQ. NPS, NKE, TTQ, GKQ,TAQ, VKQ, NYQ, NTP, TEQ, SKQ, NIQ, YNQ, TVQ, GTQ, NTR, TLQ, TMQ, KNQ, YTQ, NKS, NTE, NIE, TRQ, NSQ, YTK, NIK, NNK, NSK, ITK, NAK, KTK, GNQ, SNQ, HTK, DTK, NEQ, NPK, YTE, NNR, INQ, NHQ, YIQ, HNQ, ITQ, STQ, NSN, NQQ, NNP, ITE, NTN, FSQ, NNH, NMQ, NTS, NLQ, SMQ. NCQ, or VTQ15. The AAV particle of any one of embodiments 1-14, wherein [Nl] is or comprises TNTQ (SEQ ID NO: 4688), TNTK (SEQ ID NO: 4689), TNNQ (SEQ ID NO: 4690), SNNQ (SEQ ID NO: 4691), TNKQ (SEQ ID NO: 4692). TNNE (SEQ ID NO: 4693), SNKQ (SEQ ID NO: 4694), SNTQ (SEQ ID NO: 4695), TTNQ (SEQ ID NO: 4696), TNDQ (SEQ ID NO: 4697), TTIQ (SEQ ID NO: 4698), RNTQ (SEQ ID NO: 4699), TTKQ (SEQ ID NO: 4700), TTSQ (SEQ ID NO: 4701), TTDQ (SEQ ID NO: 4702), TNPS (SEQ ID NO: 4703), TNKE (SEQ ID NO: 4704). TTTQ (SEQ ID NO: 4705), TGKQ (SEQ ID NO: 4706), TTAQ (SEQ ID NO: 4707), TVKQ (SEQ ID NO: 4708). TNYQ (SEQ ID NO: 4709), TNTP (SEQ ID NO: 4710). STKQ (SEQ ID NO: 4711), TTEQ (SEQ ID NO: 4712). TSKQ (SEQ ID NO: 4713), TNIQ (SEQ ID NO: 4714), TYNQ (SEQ ID NO: 4715), STIQ (SEQ ID NO: 4716), TTVQ (SEQ ID NO: 4717), TGTQ (SEQ ID NO: 4718), TNTR (SEQ ID NO: 4719). TTLQ (SEQ ID NO: 4720), TTMQ (SEQ ID NO: 4721), ANNQ (SEQ ID NO: 4722), SNIQ (SEQ ID NO: 4723), TKNQ (SEQ ID NO: 4724), TYTQ (SEQ ID NO: 4725). TNKS (SEQ ID NO: 4726), SNTE (SEQ ID NO: 4727), TNTE (SEQ ID NO: 4728), TNIE (SEQ ID NO: 4729). TTRQ (SEQ ID NO: 4730), TNSQ (SEQ ID NO: 4731). TYTK (SEQ ID NO: 4732), TTTK (SEQ ID NO: 4733), TNIK (SEQ ID NO: 4734). SNTK (SEQ ID NO: 4735), TNNK (SEQ ID NO: 4736), TNSK (SEQ ID NO: 4737), TSTK (SEQ ID NO: 4738). TITK (SEQ ID NO: 4739), INTK (SEQ ID NO: 4740), TNAK (SEQ ID NO: 4741), TKTK (SEQ ID NO: 4742), STNQ (SEQ ID NO: 4743), ANTK (SEQ ID NO: 4744), RNNQ (SEQ ID NO: 4745), TGNQ (SEQ ID NO: 4746), TSNQ (SEQ ID NO: 4747), THTK (SEQ ID NO: 4748), TDTK (SEQ ID NO: 4749), TNEQ (SEQ ID NO: 4750), CNTQ (SEQ ID NO: 4751), TNPK (SEQ ID NO: 4752), INNQ (SEQ ID NO: 4753), TYTE (SEQ ID NO: 4754), NNNQ (SEQ ID NO: 4755), KNNQ (SEQ ID NO: 4756), TNNR (SEQ ID NO: 4757), LNNQ (SEQ ID NO: 4758), TINQ (SEQ ID NO: 4759), TNHQ (SEQ ID NO: 4760), STTQ (SEQ ID NO: 4761), SNSQ (SEQ ID NO: 4762), STSQ (SEQ ID NO: 4763), TYIQ (SEQ ID NO: 4764). SGTQ (SEQ ID NO: 4765), THNQ (SEQ ID NO: 4766), TITQ (SEQ ID NO: 4767), TSTQ (SEQ ID NO: 4768), TNSN (SEQ ID NO: 4769), TNQQ (SEQ ID NO: 4770), RNIQ (SEQ ID NO: 4771), TNNP (SEQ ID NO: 4772), TITE (SEQ ID NO: 4773), TNTN (SEQ ID NO: 4774), TFSQ (SEQ ID NO: 4775), RNSQ (SEQ ID NO: 4776), INTQ (SEQ ID NO: 4777). RNTE (SEQ ID NO: 4778), TNNH (SEQ ID NO: 4779), TNMQ (SEQ ID NO: 4780), RTTQ (SEQ ID NO: 4781), SNIE (SEQ ID NO: 4782). TNTS (SEQ ID NO: 4783), KNTQ (SEQ ID NO: 4784), TNLQ (SEQ ID NO: 4785), TSMQ (SEQ ID NO: 4786). SYTQ (SEQ ID NO: 4787), TNCQ (SEQ ID NO: 4788), SSTQ (SEQ ID NO: 4789). TVTQ (SEQ ID NO: 4790), or QNTQ (SEQ ID NO: 4791).16. The AAV particle of embodiment 15. wherein [Nl] is or comprises TNTQ (SEQ ID NO: 4688).17. The AAV particle of embodiment 15, wherein [Nl] is or comprises TNTK (SEQ ID NO: 4689).18. The AAV particle of any one of embodiments 1-17, wherein [N1]-[N2] comprises:(i) TQDWHR (SEQ ID NO: 4686), TKDWHR (SEQ ID NO: 4792), NQDWHR (SEQ ID NO: 4793), KQDWHR (SEQ ID NO: 4794). NEDWHR (SEQ ID NO: 4795), DQDWHR (SEQ ID NO: 4796), IQDWHR (SEQ ID NO: 4797), SQDWHR (SEQ ID NO: 4798), PSDWHR (SEQ ID NO: 4799), KEDWHR (SEQ ID NO: 4800), AQDWHR (SEQ ID NO: 4801), YQDWHR (SEQ ID NO: 4802). TPDWHR (SEQ ID NO: 4803), EQDWHR (SEQ ID NO: 4804). VQDWHR (SEQ ID NO: 4805), TRDWHR (SEQ ID NO: 4806), LQDWHR (SEQ ID NO: 4807), MQDWHR (SEQ ID NO: 4808). KSDWHR (SEQ ID NO: 4809), TEDWHR (SEQ ID NO: 4810). IEDWHR (SEQ ID NO: 4811), RQDWHR (SEQ ID NO: 4812), IKDWHR (SEQ ID NO: 4813). NKDWHR (SEQ ID NO: 4814), SKDWHR (SEQ ID NO: 4815), AKDWHR (SEQ ID NO: 4816), PKDWHR (SEQ ID NO: 4817). NRDWHR (SEQ ID NO: 4818), HQDWHR (SEQ ID NO: 4819), SNDWHR (SEQ ID NO: 4820), QQDWHR (SEQ ID NO: 4821), NPDWHR (SEQ ID NO: 4822). TNDWHR (SEQ ID NO: 4823), NHDWHR (SEQ ID NO: 4824), TSDWHR (SEQ ID NO: 4825), or CQDWHR (SEQ ID NO: 4826);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i). e.g., any 2, 3, 4. or 5 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, tw o. or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).19. The AAV particle of any one of embodiments 1-18, wherein [N1]-[N2] comprises:(i) NTQDWHR (SEQ ID NO: 4827), NTKDWHR (SEQ ID NO: 4828), NNQDWHR (SEQ ID NO: 4829), NKQDWHR (SEQ ID NO: 4830), NNEDWHR (SEQ ID NO: 4831), TNQDWHR (SEQ ID NO: 4832), NDQDWHR (SEQ ID NO: 4833), TIQDWHR (SEQ ID NO: 4834), TKQDWHR (SEQ ID NO: 4835), TSQDWHR (SEQ ID NO: 4836), TDQDWHR (SEQ ID NO: 4837), NPSDWHR (SEQ ID NO: 4838), NKEDWHR (SEQ ID NO: 4839). TTQDWHR (SEQ ID NO: 4840), GKQDWHR (SEQ ID NO: 4841), TAQDWHR (SEQ ID NO: 4842). VKQDWHR (SEQ ID NO: 4843), NYQDWHR (SEQ ID NO: 4844), NTPDWHR (SEQ ID NO: 4845), TEQDWHR (SEQ ID NO: 4846), SKQDWHR (SEQ ID NO: 4847), NIQDWHR (SEQ ID NO: 4848), YNQDWHR (SEQ ID NO: 4849), TVQDWHR (SEQ ID NO: 4850), GTQDWHR (SEQ ID NO: 4851). NTRDWHR (SEQ ID NO: 4852), TLQDWHR (SEQ ID NO: 4853), TMQDWHR (SEQ ID NO: 4854), KNQDWHR (SEQ ID NO: 4855). YTQDWHR (SEQ ID NO: 4856), NKSDWHR (SEQ ID NO: 4857). NTEDWHR (SEQ ID NO: 4858), NIEDWHR (SEQ ID NO: 4859), TRQDWHR (SEQ ID NO: 4860). NSQDWHR (SEQ ID NO: 4861), YTKDWHR (SEQ ID NO: 4862), TTKDWHR (SEQ ID NO: 4863). NIKDWHR (SEQ ID NO: 4864). NNKDWHR (SEQ IDNO: 4865), NSKDWHR (SEQ ID NO: 4866), STKDWHR (SEQ ID NO: 4867), ITKDWHR (SEQ ID NO: 4868), NAKDWHR (SEQ ID NO: 4869), KTKDWHR (SEQ ID NO: 4870), GNQDWHR (SEQ ID NO: 4871), SNQDWHR (SEQ ID NO: 4872), HTKDWHR (SEQ ID NO: 4873), DTKDWHR (SEQ ID NO: 4874), NEQDWHR (SEQ ID NO: 4875), NPKDWHR (SEQ ID NO: 4876), YTEDWHR (SEQ ID NO: 4877), NNRDWHR (SEQ ID NO: 4878), INQDWHR (SEQ ID NO: 4879), NHQDWHR (SEQ ID NO: 4880), YIQDWHR (SEQ ID NO: 4881), HNQDWHR (SEQ ID NO: 4882), ITQDWHR (SEQ ID NO: 4883), STQDWHR (SEQ ID NO: 4884), NSNDWHR (SEQ ID NO: 4885), NQQDWHR (SEQ ID NO: 4886), NNPDWHR (SEQ ID NO: 4887). ITEDWHR (SEQ ID NO: 4888). NTNDWHR (SEQ ID NO: 4889), FSQDWHR (SEQ ID NO: 4890), NNHDWHR (SEQ ID NO: 4891), NMQDWHR (SEQ ID NO: 4892), NTSDWHR (SEQ ID NO: 4893). NLQDWHR (SEQ ID NO: 4894), SMQDWHR (SEQ ID NO: 4895), NCQDWHR (SEQ ID NO: 4896), or VTQDWHR (SEQ ID NO: 4897);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5, or 6 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one. two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).20. The AAV particle of any one of embodiments 1-19, wherein [N1]-[N2] is or comprises:(i) TNTQDWHR (SEQ ID NO: 4898), TNTKDWHR (SEQ ID NO: 4899), TNNQDWHR (SEQ ID NO: 4900), SNNQDWHR (SEQ ID NO: 4901), TNKQDWHR (SEQ ID NO: 4902), TNNEDWHR (SEQ ID NO: 4903), SNKQDWHR (SEQ ID NO: 4904), SNTQDWHR (SEQ ID NO: 4905), TTNQDWHR (SEQ ID NO: 4906), TNDQDWHR (SEQ ID NO: 4907), TTIQDWHR (SEQ ID NO: 4908), RNTQDWHR (SEQ ID NO: 4909), TTKQDWHR (SEQ ID NO: 4910), TTSQDWHR (SEQ ID NO: 4911), TTDQDWHR (SEQ ID NO: 4912), TNPSDWHR (SEQ ID NO: 4913), TNKEDWHR (SEQ ID NO: 4914), TTTQDWHR (SEQ ID NO: 4915), TGKQDWHR (SEQ ID NO: 4916), TTAQDWHR (SEQ ID NO: 4917), TVKQDWHR (SEQ ID NO: 4918), TNYQDWHR (SEQ ID NO: 4919), TNTPDWHR (SEQ ID NO: 4920), STKQDWHR (SEQ ID NO: 4921). TTEQDWHR (SEQ ID NO: 4922), TSKQDWHR (SEQ ID NO: 4923), TNIQDWHR (SEQ ID NO: 4924), TYNQDWHR (SEQ ID NO: 4925), STIQDWHR (SEQ ID NO: 4926), TTVQDWHR (SEQ ID NO: 4927), TGTQDWHR (SEQ ID NO: 4928), TNTRDWHR (SEQ ID NO: 4929), TTLQDWHR (SEQ ID NO: 4930), TTMQDWHR (SEQ ID NO: 4931). ANNQDWHR (SEQ ID NO: 4932), SNIQDWHR (SEQ ID NO: 4933), TKNQDWHR (SEQ ID NO: 4934), TYTQDWHR (SEQ ID NO: 4935), TNKSDWHR (SEQ ID NO: 4936), SNTEDWHR (SEQ ID NO: 4937), TNTEDWHR (SEQ ID NO: 4938), TNIEDWHR (SEQ ID NO: 4939), TTRQDWHR (SEQ ID NO: 4940), TNSQDWHR (SEQ ID NO: 4941). TYTKDWHR (SEQ ID NO: 4942), TTTKDWHR (SEQ ID NO: 4943), TNIKDWHR (SEQ ID NO: 4944), SNTKDWHR (SEQ ID NO: 4945). TNNKDWHR (SEQ ID NO: 4946), TNSKDWHR (SEQ ID NO: 4947),TSTKDWHR (SEQ ID NO: 4948), TITKDWHR (SEQ ID NO: 4949), INTKDWHR (SEQ ID NO: 4950), TNAKDWHR (SEQ ID NO: 4951), TKTKDWHR (SEQ ID NO: 4952), STNQDWHR (SEQ ID NO: 4953), ANTKDWHR (SEQ ID NO: 4954), RNNQDWHR (SEQ ID NO: 4955), TGNQDWHR (SEQ ID NO: 4956). TSNQDWHR (SEQ ID NO: 4957), THTKDWHR (SEQ ID NO: 4958), TDTKDWHR (SEQ ID NO: 4959), TNEQDWHR (SEQ ID NO: 4960), CNTQDWHR (SEQ ID NO: 4961), TNPKDWHR (SEQ ID NO: 4962), INNQDWHR (SEQ ID NO: 4963), TYTEDWHR (SEQ ID NO: 4964), NNNQDWHR (SEQ ID NO: 4965). KNNQDWHR (SEQ ID NO: 4966), TNNRDWHR (SEQ ID NO: 4967). LNNQDWHR (SEQ ID NO: 4968), TINQDWHR (SEQ ID NO: 4969), TNHQDWHR (SEQ ID NO: 4970), STTQDWHR (SEQ ID NO: 4971), SNSQDWHR (SEQ ID NO: 4972), STSQDWHR (SEQ ID NO: 4973), TYIQDWHR (SEQ ID NO: 4974). SGTQDWHR (SEQ ID NO: 4975), THNQDWHR (SEQ ID NO: 4976). TITQDWHR (SEQ ID NO: 4977), TSTQDWHR (SEQ ID NO: 4978), TNSNDWHR (SEQ ID NO: 4979), TNQQDWHR (SEQ ID NO: 4980), RN1QDWHR (SEQ ID NO: 4981). TNNPDWHR (SEQ ID NO: 4982), TITEDWHR (SEQ ID NO: 4983). TNTNDWHR (SEQ ID NO: 4984), TFSQDWHR (SEQ ID NO: 4985), RNSQDWHR (SEQ ID NO: 4986). INTQDWHR (SEQ ID NO: 4987). RNTEDWHR (SEQ ID NO: 4988). TNNHDWHR (SEQ ID NO: 4989), TNMQDWHR (SEQ ID NO: 4990). RTTQDWHR (SEQ ID NO: 4991). SNIEDWHR (SEQ ID NO: 4992), TNTSDWHR (SEQ ID NO: 4993). KNTQDWHR (SEQ ID NO: 4994), TNLQDWHR (SEQ ID NO: 4995), TSMQDWHR (SEQ ID NO: 4996), SYTQDWHR (SEQ ID NO: 4997), TNCQDWHR (SEQ ID NO: 4998). SSTQDWHR (SEQ ID NO: 4999). TVTQDWHR (SEQ ID NO: 5000). or QNTQDWHR (SEQ ID NO: 5001);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i). e.g., any 2, 3, 4, 5, 6, or 7 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions, relative to any one of the amino acid sequences in (i).21. The AAV particle of embodiment 20, wherein [N1]-[N2] is or comprises TNTQDWHR (SEQ ID NO: 4898).22. The AAV particle of embodiment 20, wherein [N1]-[N2] is or comprises TNTKDWHR (SEQ ID NO: 4899).23. The AAV particle of any one of embodiments 1-22, wherein the AAV capsid variant comprises one, two, or all of: an amino acid other than Q at position 590 (e.g.. I, V, T, M, S, N. L, or F), an amino acid other than A at position 591 (e.g., Y, P. N, S, T. G, E, V. W. F. Q). and / or an amino acid other than Q at position 592 (e.g.. G, N. K, H, R. E, L, P. or M), as numbered according to SEQ ID NO: 138 or 981.24. The AAV particle of any one of embodiments 1-23, wherein the AAV capsid variant comprises an amino acid other than Q at position 590 (e.g., I, V, T, M, S, N, L, or F), as numbered according to SEQ ID NO: 138 or 981.25. The AAV particle of any one of embodiments 1-24, wherein the AAV capsid variant comprises the amino acid I at position 590, as numbered according to SEQ ID NO: 138 or 981.26. The AAV particle of any one of embodiments 1-24, wherein the AAV capsid variant comprises the amino acid V at position 590, as numbered according to SEQ ID NO: 138 or 981. 1. The AAV particle of any one of embodiments 1-26, wherein the AAV capsid variant comprises the amino acid A at position 591 and / or the amino acid Q at position 592, as numbered according to SEQ ID NO: 138 or 981.28. The AAV particle of any one of embodiments 1-27, wherein [N3] comprises amino acids Xs, Xe, and X?, wherein Xs is I, V, T. M, S, N, L, or F.29. The AAV particle of any one of embodiments 1-28, wherein Xs is I or V.30. The AAV particle of embodiment 29, wherein Xs is I.31. The AAV particle of any one of embodiments 1-30, wherein:(i) X6is A, Y, P, N, S, T, G, E, V. W, F, or Q; and / or(ii) X7is Q, G, N, K, H, R, E, L, P, or M.32. The AAV particle of any one of embodiments 1-31, wherein [N3] comprises IA, IY, VP, IN, VN, VY, VA, IS, IT, TA, MA, SA, IG, IE, IV, NA, LA, IP, FA, VS, VT, IW, IF, IQ, VQ, AQ, AG, YQ, PQ, AN, NQ, SG, SQ, TQ, GQ, EQ, AK, AH, AR, AE, AL, AP, TM, SM, WQ, FQ, QQ, FM, AM. or SN.33. The AAV particle of any one of embodiments 1-32, wherein [N3] is or comprises IAQ, IAG. IYQ, VPQ, IAN, INQ, VNQ, VYQ, VAN, ISG, ISQ, VAQ, ITQ, TAQ, MAQ, SAQ, IGQ, IEQ. IVQ, NAQ, LAQ. IAK, IAH, IPQ, IAR, IAE. IAL, IAP. FAQ. VSQ. VTM, ISM. IWQ, IFQ, IQQ. VQQ, IFM. IAM, or ISN.34. The AAV particle of any one of embodiments 1-33, wherein [N3] is or comprises IAQ.35. The AAV particle of any one of embodiments 1-34, wherein [N2]-[N3] comprises:(i) DWHRIA (SEQ ID NO: 5002), DWHRIY (SEQ ID NO: 5003), DWHRVP (SEQ ID NO: 5004), DWHRIN (SEQ ID NO: 5005), DWHRVN (SEQ ID NO: 5006), DWHRVY (SEQ ID NO: 5007), DWHRVA (SEQ ID NO: 5008), DWHRIS (SEQ ID NO: 5009), DWHRIT (SEQ ID NO: 5010), DWHRTA (SEQ ID NO: 5011), DWHRMA (SEQ ID NO: 5012), DWHRSA (SEQ ID NO: 5013), DWHRIG (SEQ ID NO: 5014), DWHRIE (SEQ ID NO: 5015), DWHRIV (SEQ ID NO: 5016), DWHRNA (SEQ ID NO: 5017), DWHRLA (SEQ ID NO: 5018), DWHRIP (SEQ ID NO: 5019), DWHRFA (SEQ ID NO: 5020), DWHRVS (SEQ ID NO: 5021). DWHRVT (SEQ ID NO: 5022), DWHRIW (SEQ ID NO: 5023), DWHRIF (SEQ ID NO: 5024), DWHRIQ (SEQ ID NO: 5025). or DWHRVQ (SEQ ID NO: 5026);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, or 5 amino acids, e.g.. consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two. or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one. two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).36. The AAV particle of any one of embodiments 1-35, wherein [N2]-[N3] is or comprises:(i) DWHRIAQ (SEQ ID NO: 5027), DWHRIAG (SEQ ID NO: 5028), DWHRIYQ (SEQ ID NO: 5029), DWHRVPQ (SEQ ID NO: 5030), DWHRIAN (SEQ ID NO: 5031). DWHRINQ (SEQ ID NO: 5032), DWHRVNQ (SEQ ID NO: 5033). DWHRVYQ (SEQ ID NO: 5034), DWHRVAN (SEQ ID NO: 5035), DWHRISG (SEQ ID NO: 5036), DWHRISQ (SEQ ID NO: 5037), DWHRVAQ (SEQ ID NO: 5038), DWHRITQ (SEQ ID NO: 5039), DWHRTAQ (SEQ ID NO: 5040), DWHRMAQ (SEQ ID NO: 5041), DWHRSAQ (SEQ ID NO: 5042), DWHRIGQ (SEQ ID NO: 5043), DWHRIEQ (SEQ ID NO: 5044), DWHRIVQ (SEQ ID NO: 5045), DWHRNAQ (SEQ ID NO: 5046), DWHRLAQ (SEQ ID NO: 5047), DWHRIAK (SEQ ID NO: 5048), DWHRIAH (SEQ ID NO: 5049), DWHRIPQ (SEQ ID NO: 5050), DWHRIAR (SEQ ID NO: 5051), DWHRIAE (SEQ ID NO: 5052), DWHRIAL (SEQ ID NO: 5053), DWHRIAP (SEQ ID NO: 5054), DWHRFAQ (SEQ ID NO: 5055), DWHRVSQ (SEQ ID NO: 5056), DWHRVTM (SEQ ID NO: 5057), DWHRISM (SEQ ID NO: 5058), DWHRIWQ (SEQ ID NO: 5059), DWHRIFQ (SEQ ID NO: 5060), DWHRIQQ (SEQ ID NO: 5061), DWHRVQQ (SEQ ID NO: 5062), DWHRIFM (SEQ ID NO: 5063), DWHRIAM (SEQ ID NO: 5064), or DWHRISN (SEQ ID NO: 5065);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5, or 6 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).37. The AAV particle of any one of embodiments 1-36, wherein [N2]-[N3] is or comprises DWHRIAQ (SEQ ID NO: 5027).38. The AAV particle of any one of embodiments 1-37, wherein [N1]-[N2]-[N3] is or comprises:(i) the amino acid sequence of any one of SEQ ID NOs: 343-538:(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5, 6, 7, 8. 9, or 10 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).39. The AAV particle of embodiment 38, wherein [N1]-[N2]-[N3] is or comprises TNTQDWHRIAQ (SEQ ID NO: 343).40. The AAV particle of embodiment 38, wherein [N1]-[N2]-[N3] is or comprises TNTKDWHRIAQ (SEQ ID NO: 344).41. The AAV particle of any one of embodiments 1-40, wherein the AAV capsid variant comprises one, two, three, or all of: an amino acid other than T at position 593 (e.g., S, N, P, A, or I), an amino acid other than G at position 594 (e.g.. N, D, R, V, A, S, or Q), an amino acid other than W at position 595 (e.g., S, C, R, L, or G), and / or an amino acid other than V at position 596 (e.g., A, S, I, C, G, D, F, L, or T), as numbered according to SEQ ID NO: 138 or 981.42. The AAV particle of any one of embodiments 1-40, wherein the AAV capsid variant comprises tire amino acid T at position 593. the amino acid G at position 594. the amino acid W at position 595, and the amino acid V at position 596, as numbered according to SEQ ID NO: 138 or 981.43. The AAV particle of any one of embodiments 1-42, wherein the AAV capsid variant further comprises [N4], wherein [N4] comprises amino acids X8, X9, Xio, and Xu, wherein:(i) Xsis T, S, N, P. A, or I;(ii) X9is G, N. D, R. V, A, S. or Q;(iii) X10is W, S, C. R, L. or G; and / or(iv) Xnis V, A, S. I. C, G. D, F, L. or T.44. The AAV particle of embodiment 43, wherein [N4] comprises TG, TN, SN, NN, SG, PG, TD, AG, IG, NG, TR, TV, TA, TS, SV, TQ, WV, WA, WS, WI, WC, WG, CV, RV, LV, GV, WD, WF, WL, WT, GW, NW, GS, DW, GC, GR, GL, GG, RW, VW, AW, SW, or QW.45. The AAV particle of embodiment 43 or 44, wherein [N4] comprises TGW, TNW, SNW. NNW, SGW, PGW, TGS, TDW, TGC, TGR, TGL, TGG, AGW, IGW, NGW. TRW, TVW, TAW, TSW. SVW, TQW, GWV, GW A. NWS, NWV, NWI, GWS, GWI, GWC, GWG, GSV, DWV. GCV. GRV, GLV, GGV. GWD, GWF, RWV. VWV, GWL, AWV, SWV, GWT, or QWV.46. The AAV particle of any one of embodiments 43-45, wherein [N4] is or comprises TGWV (SEQ ID NO: 5066), TGWA (SEQ ID NO: 5067), TNWS (SEQ ID NO: 5068), SNWV (SEQ ID NO: 5069), TNWV (SEQ ID NO: 5070). TNWI (SEQ ID NO: 5071), NNWV (SEQ ID NO: 5072), TGWS (SEQ ID NO: 5073), TGWI (SEQ ID NO: 5074), TGWC (SEQ ID NO: 5075), TGWG (SEQ ID NO: 5076). SGWV (SEQ ID NO: 5077), PGWV (SEQ ID NO: 5078), TGSV (SEQ ID NO: 5079). TDWV (SEQ ID NO: 5080), TGCV (SEQ ID NO: 5081). TGRV (SEQ ID NO: 5082), TGLV (SEQ ID NO: 5083), TGGV (SEQ ID NO: 5084), AGWV (SEQ ID NO: 5085). IGWV (SEQ ID NO: 5086), TGWD (SEQ ID NO: 5087). NGWV (SEQ ID NO: 5088), TGWF (SEQ ID NO: 5089). TRWV (SEQ ID NO: 5090), TVWV (SEQ ID NO: 5091), TGWL (SEQ ID NO: 5092), TAWV (SEQ ID NO: 5093), TSWV (SEQ ID NO: 5094). TGWT (SEQ ID NO: 5095), SVWV (SEQ ID NO: 5096), TQWV (SEQ ID NO: 5097), or PGWG (SEQ ID NO: 5098).47. The AAV particle of any one of embodiments 43-46, wherein [N4] is or comprises TGWV (SEQ ID NO: 5066).48. The AAV particle of any one of embodiments 43-47, wherein [N1]-[N2]-[N3]-[N4] is or comprises:(i) the amino acid sequence of any one of SEQ ID NOs: 201-245, 247-250, 253-255, 257-265, 268-274, 276-286, 288, 290-297, 299-303, 305-309, 311, 313-319, 323-328, 330-337, 339-342, 539-542, 544, 546, 547, 549-557, 559-589. 592, 593, 595, 596, 598, 599, 601-608, 610-614, 616-622, 625, 628, 630, 631, 633. 636, 638, 639-646, 649, 651-657, 667, 669, 670, 672, 673, 679-683, 685-690. 692, 693, 695, 697, 699-701, 703-705, 708-710, 712-717, 719-723, 728-731, 733-738, 740, and 742;(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5, 6, 7, 8. 9, 10. 11, 12, or 13 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).49. The AAV particle of embodiment 48, wherein [N1]-[N2]-[N3]-[N4] is or comprises TNTQDWHRIAQTGWV (SEQ ID NO: 201).50. The AAV particle of any one of embodiment 48. wherein [N1]-[N2]-[N3]-[N4] is or comprises TNTKDWHRIAQTGWV (SEQ ID NO: 202).51. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising an amino acid sequence having the following fonnula: [N1]-[N2]-[N3] (SEQ ID NO: 4683), wherein [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682) and wherein:(i) LN 1 J comprises amino acids Xi, X2, X3, and X4. wherein X4is Q, P, or a conservative substitution thereof; and / or(ii) [N3] amino acids X5, X«, and X?, wherein Xs is I. V, or a conservative substitution thereof.52. The AAV particle of embodiment 51, wherein the AAV capsid variant comprises the amino acid Q at position 585. as numbered according to SEQ ID NO: 138 or 981.53. The AAV particle of embodiment 51 or 52, wherein the AAV capsid variant comprises one, tw o, three, or all of:(i) an amino acid other than T at position 582 (e.g.. S);(ii) an amino acid other than N at position 583 (e.g., T, G, S, I, or V);(iii) an amino acid other than H at position 584 (e.g., N, I, S, A. V, or L); and / or(iv) an amino acid other than Q at position 585 (e.g., P), wherein the amino acids arc as numbered according to SEQ ID NO: 138 or 981.54. The AAV particle of any one of embodiments 51-53, wherein [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4is Q or P.55. The AAV particle of any one of embodiments 51-54, wherein X4is Q.56. The AAV particle of any one of embodiments 51-55, wherein the AAV capsid variant comprises an amino acid other than H at position 584 (e.g., T), as numbered according to SEQ ID NO: 138 or 981.57. The AAV particle of any one of embodiments 51-56, wherein the AAV capsid variant comprises the amino acid T at position 584, as numbered according to SEQ ID NO: 138 or 981.58. The AAV particle of any one of embodiments 51-57, wherein:(i) Xi is T or S;(ii) X2 is N, T, G, S, I, or V; and / or(iii) X3is T, N, I, S, A, V, or L.59. The AAV particle of any one of embodiments 51-58, wherein [Nl] comprises TN, TT. TG, ST, TS, TI, TV, TQ, NQ, IQ. SQ, AQ, VQ, TP, LQ, NT, TA, NI. GT, IT. NN, TL, NS, or VT.60. The AAV particle of any one of embodiments 51-59, wherein [Nl] comprises TNT, TTN, TTI. TTS, TTT, TTA, TNI. TTV. TGT. STT, TST, TIT, TNN, TTL, TNS, TVT, NTQ, TNQ, TIQ, TSQ, TTQ, TAQ. NIQ, TVQ, GTQ, STQ, ITQ. NTP. NNQ. TLQ. NSQ. or VTQ.61. The AAV particle of any one of embodiments 51-60, wherein [Nl] is or comprises TNTQ (SEQ ID NO: 4688), TTNQ (SEQ ID NO: 4696), TTIQ (SEQ ID NO: 4698), TTSQ (SEQ ID NO: 4701), TTTQ (SEQ ID NO: 4705), TTAQ (SEQ ID NO: 4707), TNIQ (SEQ ID NO: 4714), TTVQ (SEQ ID NO: 4717). TGTQ (SEQ ID NO: 4718), STTQ (SEQ ID NO: 4761). TSTQ (SEQ ID NO: 4768), TITQ (SEQ ID NO: 4767), TNTP (SEQ ID NO: 4710), TNNQ (SEQ ID NO: 4690). TTLQ (SEQ ID NO: 4720), TNSQ (SEQ ID NO: 4731), or TVTQ (SEQ ID NO: 4790).62. The AAV particle of embodiment 61, wherein [Nl] is or comprises TNTQ (SEQ ID NO: 4688).63. The AAV particle of any one of embodiments 51-62, wherein [N1]-[N2] comprises:(i) TQDWHR (SEQ ID NO: 4686), NQDWHR (SEQ ID NO: 4793), IQDWHR (SEQ ID NO: 4797), SQDWHR (SEQ ID NO: 4798), AQDWHR (SEQ ID NO: 4801), VQDWHR (SEQ ID NO: 4805), TPDWHR (SEQ ID NO: 4803), or LQDWHR (SEQ ID NO: 4807);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3, 4, or 5 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).64. The AAV particle of any one of embodiments 51-63, wherein [N1]-[N2] comprises:(i) NTQDWHR (SEQ ID NO: 4827), TNQDWHR (SEQ ID NO: 4832), TIQDWHR (SEQ ID NO: 4834), TSQDWHR (SEQ ID NO: 4836). TTQDWHR (SEQ ID NO: 4840), TAQDWHR (SEQ ID NO: 4842), NIQDWHR (SEQ ID NO: 4848). TVQDWHR (SEQ ID NO: 4850), GTQDWHR (SEQ ID NO: 4851), STQDWHR (SEQ ID NO: 4884). ITQDWHR (SEQ ID NO: 4883). NTPDWHR (SEQ IDNO: 4845), NNQDWHR (SEQ ID NO: 4829), TLQDWHR (SEQ ID NO: 4853), NSQDWHR (SEQ ID NO: 4861), VTQDWHR (SEQ ID NO: 4897);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5 or 6 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).65. The AAV particle of any one of embodiments 51-64, wherein [N1]-[N2] is or comprises:(i) TNTQDWHR (SEQ ID NO: 4898), TTNQDWHR (SEQ ID NO: 4906), TTIQDWHR (SEQ ID NO: 4908), TTSQDWHR (SEQ ID NO: 4911), TTTQDWHR (SEQ ID NO: 4915), TTAQDWHR (SEQ ID NO: 4917). TNIQDWHR (SEQ ID NO: 4924). TTVQDWHR (SEQ ID NO: 4927), TGTQDWHR (SEQ ID NO: 4928), STTQDWHR (SEQ ID NO: 4971), TSTQDWHR (SEQ ID NO: 4978). TITQDWHR (SEQ ID NO: 4977), TNTPDWHR (SEQ ID NO: 4920), TNNQDWHR (SEQ ID NO: 4900), TTLQDWHR (SEQ ID NO: 4930), TNSQDWHR (SEQ ID NO: 4941), TVTQDWHR (SEQ ID NO: 5000);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i). e.g., any 2, 3, 4. 5 6, or 7 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, tw o. or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).66. The AAV particle of embodiment 65, wherein [N1]-[N2] is or comprises TNTQDWHR (SEQ ID NO: 4898).67. The AAV particle of any one of embodiments 51-66, w herein the AAV capsid variant comprises one. two, or all of: an amino acid other than Q at position 590 (e.g., I or V), an amino acid other than A at position 591 (e.g., P. S, Y, or N), and / or an amino acid other than Q at position 592 (e g., G or N), as numbered according to SEQ ID NO: 138 or 981.68. The AAV particle of any one of embodiments 51-67, wherein the AAV capsid variant comprises an amino acid other than Q at position 590 (e.g., I or V), as numbered according to SEQ ID NO: 138 or 981.69. The AAV particle of any one of embodiments 51-68, wherein the AAV capsid variant comprises the amino acid I at position 590. as numbered according to SEQ ID NO: 138 or 981.70. The AAV particle of any one of embodiments 51-68, wherein the AAV capsid variant comprises the amino acid V at position 590, as numbered according to SEQ ID NO: 138 or 981.71. The AAV particle of any one of embodiments 51-70, wherein the AAV capsid variant comprises the amino acid A at position 591 and / or the amino acid Q at position 592, as numbered according to SEQ ID NO: 138 or 981.72. The AAV particle of any one of embodiments 51-71. wherein [N3] comprises amino acids X5, X6, and X7, wherein X5is I or V.73. The AAV particle of embodiment 72. wherein Xs is I.74. The AAV particle of any one of embodiments 51-73, wherein:(i) X6is A. P. S, Y. or N; and / or(ii) X? is Q, G, or N.75. The AAV particle of any one of embodiments 51-74, wherein [N3] comprises IA, VP, VA, VS. IY, IN, IS, AQ, AG, PQ, SQ. AN, YQ, or NQ.76. The AAV particle of any one of embodiments 51-75, wherein [N3] is or comprises IAQ, IAG, VPQ, VAQ, VSQ, IAN, IYQ, INQ, or ISQ.77. The AAV particle of any one of embodiments 51-76, wherein [N3] is or comprises IAQ.78. The AAV particle of any one of embodiments 51-77, wherein [N2]-[N3] comprises:(i) DWHRIA (SEQ ID NO: 5002), DWHRVP (SEQ ID NO: 5004), DWHRVA (SEQ ID NO: 5008), DWHRVS (SEQ ID NO: 5021), DWHRIY (SEQ ID NO: 5003), DWHRIN (SEQ ID NO: 5005), or DWHRIS (SEQ ID NO: 5009);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, or 5 amino acids, e.g.. consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).79. The AAV particle of any one of embodiments 51-78, wherein [N2]-[N3] is or comprises:(i) DWHRIAQ (SEQ ID NO: 5027), DWHRIAG (SEQ ID NO: 5028), DWHRVPQ (SEQ ID NO: 5030), DWHRVAQ (SEQ ID NO: 5038), DWHRVSQ (SEQ ID NO: 5056), DWHRIAN (SEQ ID NO: 5031), DWHRIYQ (SEQ ID NO: 5029), DWHRINQ (SEQ ID NO: 5032), or DWHRISQ (SEQ ID NO: 5037);(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5, or 6 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).80. The AAV particle of any one of embodiments 51-79, wherein IN2J-[N3J is or comprises DWHRIAQ (SEQ ID NO: 5027).81. The AAV particle of any one of embodiments 51-80, wherein [N1]-[N2]-[N3] is or comprises:(i) the amino acid sequence of any one of SEQ ID NOs: 343, 350. 352, 355, 359. 361, 364, 367. 370. 371, 373, 374. 376, 377, 378, 381, 395. 420, 454, 457. 460, 464, 481. 482, 488, 493, 494, 516. 525, 536;(ii) an amino acid sequence comprising any portion of an amino acid sequence in (i). e.g., any 2, 3, 4. 5, 6, 7, 8, 9. or 10 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).82. The AAV particle of any one of embodiments 51-81, wherein [N1]-[N2]-[N3] is or comprises TNTQDWHRIAQ (SEQ ID NO: 343).83. The AAV particle of any one of embodiments 51-82, wherein the AAV capsid variant comprises the amino acid W at position 595, as numbered according to SEQ ID NO: 138 or 981.84. The AAV particle of any one of embodiments 51-83, wherein the AAV capsid variant comprises one. two or all of: an amino acid other than T (e.g.. S or N) at position 593, an amino acid other than G (e.g., N) a position 594, and / or an amino acid other than V (e.g.. A, I, or S) at position 596, as numbered according to SEQ ID NO: 138 or 981.85. The AAV particle of any one of embodiments 51-84, wherein the AAV capsid variant comprises:(i) the amino acid T at position 593. the amino acid G at position 594, the amino acid W at position 595, and the amino acid V at position 596, as numbered according to SEQ ID NO: 138 or 981;(ii) the amino acid T at position 593, the amino acid G at position 594, the amino acid W at position 595, and the amino acid A at position 596, as numbered according to SEQ ID NO: 138 or 981:(iii) the amino acid S at position 593, the amino acid N at position 594, the amino acid W at position 595, and the amino acid V at position 596, as numbered according to SEQ ID NO: 138 or 981:(iv) the amino acid N at position 593, the amino acid N at position 594, the amino acid W at position 595, and the amino acid V at position 596, as numbered according to SEQ ID NO: 138 or 981:(v) the amino acid T at position 593, the amino acid G at position 594, the amino acid W at position 595, and the amino acid I at position 596. as numbered according to SEQ ID NO: 138 or 981; or(vi) the amino acid T at position 593, the amino acid G at position 594, the amino acid W at position 595, and the amino acid S at position 596, as numbered according to SEQ ID NO: 138 or 981.86. The AAV particle of any one of embodiments 51-85, wherein the AAV capsid variant comprises the amino acid T at position 593, the amino acid G at position 594, the amino acid W at position 595. and the amino acid V at position 596. as numbered according to SEQ ID NO: 138 or 981.87. The AAV particle of any one of embodiments 51-86, wherein the AAV capsid variant further comprises [N4], wherein [N4] comprises amino acids X8, X9. Xio, and Xu, wherein Xiois W.88. The AAV particle of embodiment 87, wherein:(i) X8is T, S, or N;(ii) X9is G or N; and / or(iv) Xu is V, A, I, or S.89. The AAV particle of embodiment 87 or 88, wherein [N4] comprises TG, SN, NN, WV, WA, WI, WS, GW, or NW.90. The AAV particle of any one of embodiments 87-89, wherein [N4] comprises TGW. SNW, NNW, GWV, GW A, NWV, GWI. or GWS.91. The AAV particle of any one of embodiments 87-90. wherein [N4] is or comprises TGWV (SEQ ID NO: 5066), TGWA (SEQ ID NO: 5067), SNWV (SEQ ID NO: 5069), NNWV (SEQ ID NO: 5072), TGWI (SEQ ID NO: 5074), or TGWS (SEQ ID NO: 5073).92. The AAV particle of any one of embodiments 87-91. wherein [N4] is or comprises TGWV (SEQ ID NO: 5066).93. The AAV particle of any one of embodiments 87-92, wherein [N1]-[N2]-[N3]-[N4] is or comprises:(i) the amino acid sequence of any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291, 293, 301, 306, 307, 308, 309, 314, and 336;(ii) an ammo acid sequence comprising any portion of an amino acid sequence in (i), e.g., any 2, 3. 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 amino acids, e.g., consecutive amino acids, thereof;(iii) an amino acid sequence comprising one, two, or three but no more than four modifications relative to any one of the amino acid sequences in (i); or(iv) an amino acid sequence comprising one, two, or three but no more than four substitutions relative to any one of the amino acid sequences in (i).94. The AAV particle of any one of embodiments 87-93, wherein [N 1 J-[N2J-IN3 J-LN4J is or comprises TNTQDWHRIAQTGWV (SEQ ID NO: 201).95. The AAV particle of any one of embodiments 1-94, wherein [N1]-[N2]-[N3] is present in loop VIII. optionally wherein loop VIII comprises amino acids 580-599 as numbered according to SEQ ID NO: 138 or 981.96. The AAV particle of any one of embodiments 43-50 and 87-95. wherein [N4] is present in loop VIII, optionally wherein loop VIII comprises amino acids 580-599 as numbered according to SEQ ID NO: 138 or 981.97. The AAV particle of any one of embodiments 1-96, wherein [Nl] replaces amino acids 582-585 (e.g., T582, N583, H584, Q585) of SEQ ID NO: 138.98. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, and 85-97, wherein [Nl] corresponds to amino acids 582-585 (e.g., T582, N583, T584, Q585) of SEQ ID NO: 981.99. The AAV particle of any one of embodiments 1-98, wherein [Nl] is present at amino acids 582-585, as numbered according to SEQ ID NO: 138 or 981.100. The AAV particle of any one of embodiments 1-99, wherein [Nl] corresponds to positions 582-585 (e.g., T582, N583. T584. Q585) of SEQ ID NO: 981.101. The AAV particle of any one of embodiments 1-100, wherein [N2] replaces amino acids 586-589 (e.g., S586, A587. Q588. A589) of SEQ ID NO: 138.102. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, 85-99, and 101, wherein [N2] corresponds to amino acids 586-589 (e.g., D586, W587, H588, R589) of SEQ ID NO: 981.103. The AAV particle of any one of embodiments 1-102, wherein [N2] is present at amino acids 586- 589, as numbered according to SEQ ID NO: 138 or 981.104. The AAV particle of any one of embodiments 1-103, wherein [N1]-[N2] replaces amino acids 582- 589 (e.g., T582, N583. H584, Q585, S586, A587, Q588. A589) of SEQ ID NO: 138.105. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83. 85-99, and 101-104. wherein [N1]-[N2] corresponds to amino acids 582-589 (e.g., T582, N583, T584, Q585, D586, W587, H588, R589) of SEQ ID NO: 981.106. The AAV particle of any one of embodiments 1-105, wherein [N1]-[N2] is present at amino acids 582-589, as numbered according to SEQ ID NO: 138 or 981.107. The AAV particle of any one of embodiments 1-106, wherein [N3] replaces amino acids 590-592 (e.g., Q590, A591, and Q592) of SEQ ID NO: 138.108. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, 85-99, and 101-107, wherein [N3] corresponds to amino acids 590-592 (e.g., 1590, A591, and Q592) of SEQ ID NO: 981.109. The AAV particle of any one of embodiments 1-108, wherein [N3] is present at amino acids 590- 592, as numbered according to SEQ ID NO: 138 or 981.110. The AAV particle of any one of embodiments 1-109, wherein [N2]-[N3] replaces amino acids 586- 592 (e.g., S586, A587, Q588, A589, Q590, A591, and Q592) of SEQ ID NO: 138.111. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83, 85-99, and 101-110, wherein [N2]-[N3] corresponds to amino acids 586-592 (e.g., D586, W587, H588, R589. 1590, A591. and Q592) of SEQ ID NO: 981.112. The AAV particle of any one of embodiments 1-111, wherein [N2]-[N3] is present at amino acids 586-592, as numbered according to SEQ ID NO: 138 or 981.113. The AAV particle of any one of embodiments 1-112, wherein [N1]-[N2]-[N3] replaces amino acids 582-592 (e.g., T582, N583, H584, Q585, S586, A587, Q588, A589, Q590, A591, Q592) of SEQ ID NO: 138.114. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83, 85-99, and 101-113, wherein [N1]-[N2]-[N3] corresponds to amino acids 582-592 (e.g., T582, N583, T584. Q585, D586, W587, H588, R589, 1590. A591, Q592) of SEQ ID NO: 981.115. The AAV particle of any one of embodiments 1-114, wherein [N1]-[N2]-[N3] is present at amino acids 582-592, as numbered according to SEQ ID NO: 138 or 981.116. The AAV particle of any one of embodiments 43-50 and 87-115, wherein [N4] replaces amino acids 593-596 (e.g., T593, G594, W595, and V596) of SEQ ID NO: 138.117. The AAV particle of any one of embodiments 43-49 and 87-115. wherein [N4] corresponds to amino acids 593-596 (e g., T593, G594, W595, and V596) of SEQ ID NO: 138 or 981.11 . The AAV particle of any one of embodiments 43-50 and 87-117, wherein [N4] is present at amino acids 593-596. as numbered according to SEQ ID NO: 138 or 981.119. The AAV particle of any one of embodiments 43-50 and 87-118, wherein [N2]-[N3]-[N4] replaces amino acids 586-596 (e.g., S586, A587, Q588, A589, Q590, A591, Q592. T593, G594, W595, and V596) of SEQ ID NO: 138.120. The AAV particle of any one of embodiments 43-49 and 87-119, wherein [N2]-[N3]-[N4] corresponds to amino acids 586-596 (e.g., D586, W587, H588, R589, 1590, A591, Q592, T593, G594, W595, and V596) of SEQ ID NO: 981.121. The AAV particle of any one of embodiments 43-50 and 87-120, wherein [N2]-[N3]-[N4] is present at amino acids 586-596, as numbered according to SEQ ID NO: 138 or 981.122. The AAV particle of any one of embodiments 43-50 and 87-121, wherein [N1]-[N2]-[N3]-[N4] replaces amino acids 582-596 (e.g.. T582. N583, H584, Q585, S586, A587. Q588, A589, Q590, A591. Q592, T593, G594, W595. and V596), as numbered according to SEQ ID NO: 138.123. The AAV particle of any one of embodiments 43-49 and 87-122, wherein [N1]-[N2]-[N3]-[N4] corresponds to amino acids 582-596 (e.g., T582, N583, T584, Q585, D586, W587, H588, R589, 1590, A591, Q592, T593, G594, W595, and V596) of SEQ ID NO: 981.124. The AAV particle of any one of embodiments 43-50 and 87-123, wherein [N1]-[N2]-[N3]-[N4] is present at amino acids 582-596, as numbered according to SEQ ID NO: 138 or 981.125. The AAV particle of any one of embodiments 1-124, wherein [N2] is present immediately subsequent to [Nl],126. The AAV particle of any one of embodiments 1-125, wherein [N3] is present immediately subsequent to |N2J.127. The AAV particle of any one of embodiments 43-50 and 87-126, wherein [N4] is present immediately subsequent to [N3],128. The AAV particle of any one of embodiments 1-127, wherein the AAV capsid variant comprises, from N-terminus to C-terminus. [N1]-[N2]-[N3],129. The AAV particle of any one of embodiments 43-50 and 87-128, wherein the AAV capsid variant comprises, from N-terminus to C-terminus, [N1]-[N2]-[N3]-[N4],130. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising:(a) the amino acid sequence of any one of the sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29;(b) an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8. at least 9, at least 10, at least 11, at least 12. at least 13, or at least 14 consecutive amino acids from any one of the sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29; or(c) an amino acid sequence comprising at least one, at least two, or at least three but no more than four substitutions relative to any one of the sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29; or(d) an amino acid sequence comprising at least one, at least two, or at least three but no more than four modifications relative to the amino acid sequence of any one of the sequences provided in Table 1. 2A. 2B. 14. T1 , 28, or 29.131. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising:(a) the amino acid sequence of any one of SEQ ID NOs: 201. 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291, 293, 301, 306, 307, 308, 309, 314, and 336:(b) an amino acid sequence comprising at least 3, at least 4, at least 5. at least 6, at least 7, at least8. at least 9, at least 10, at least 11, at least 12. at least 13, or at least 14 consecutive amino acids from any one of SEQ ID NOs: 201, 205-209. 211-214, 216, 219, 220, 230. 232, 237, 238. 255, 262-265. 274, 283, 286, 290, 291. 293, 301, 306. 307, 308, 309, 314, and 336:(c) an amino acid sequence comprising at least one, at least two, or at least three but no more than four substitutions, relative to the amino acid sequence of any one of SEQ ID NOs: 201. 205-209, 211-214, 216. 219, 220, 230. 232, 237, 238. 255, 262-265. 274, 283, 286. 290, 291, 293. 301, 306. 307, 308, 309, 314. and 336; or(d) an amino acid sequence comprising at least one, at least two, or at least three but no more than four modifications relative to the amino acid sequence of any one of SEQ ID NOs: 201, 205-209, 211-214. 216, 219. 220, 230, 232. 237, 238, 255. 262-265, 274. 283, 286, 290. 291, 293, 301. 306, 307. 308. 309, 314, and 336.132. The AAV particle of embodiment 130 or 131. wherein the AAV capsid variant does not comprise at least 3, at least 4, at least 5. at least 6, at least 7, at least 8, or at least 9 consecutive amino acids from TNHQSAQAQ (SEQ ID NO: 5100), optionally wherein the TNHQSAQAQ (SEQ ID NO: 5100) corresponds to amino acids 582-592 of SEQ ID NO: 138.133. The AAV particle of any one of embodiments 130-132, wherein the AAV capsid variant does not comprise: TNH, TNHQ (SEQ ID NO: 4760), TNHQS (SEQ ID NO: 5101), TNHQSA (SEQ ID NO: 5102), TNHQSAQ (SEQ ID NO: 5103), TNHQSAQA (SEQ ID NO: 5104), TNHQSAQAQ (SEQ ID NO: 5100), NHQ, NHQS (SEQ ID NO: 5105), NHQSA (SEQ ID NO: 5106), NHQSAQ (SEQ ID NO: 5107), NHQSAQA (SEQ ID NO: 5108), NHQSAQ AQ (SEQ ID NO: 5109), HQS, HQSA (SEQ ID NO: 5110), HQSAQ (SEQ ID NO: 5111), HQSAQA (SEQ ID NO: 5112), HQSAQAQ (SEQ ID NO: 5113), QSA, QSAQ (SEQ ID NO: 5114), QSAQA (SEQ ID NO: 5115). QSAQAQ (SEQ ID NO: 5116), SAQA (SEQ ID NO: 5117). or SAQAQ (SEQ ID NO: 5118).134. The AAV particle of any one of embodiments 130-124, wherein the AAV capsid variant comprises an amino acid sequence comprising at least 3. at least 4, at least 5, at least 6, at least 7. at least 8. at least9. at least 10, at least 11. at least 12, at least 13, or at least 14 consecutive amino acids from any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219. 220, 230, 232. 237, 238. 255, 262-265, 274. 283, 286. 290, 291, 293. 301, 306. 307. 308, 309. 314, and 336.135. The AAV particle of any one of embodiments 130-134, wherein the at least 3 consecutive amino acids comprise TQD.136. The AAV particle of any one of embodiments 130-135, wherein the at least 4 consecutive amino acids comprise TQDW (SEQ ID NO: 4684).137. The AAV particle of any one of embodiments 130-136, wherein the at least 5 consecutive amino acids comprise TQDWH (SEQ ID NO: 4685).138. The AAV particle of any one of embodiments 130-137. wherein the at least 6 consecutive amino acids comprise TQDWHR (SEQ ID NO: 4686).139. The AAV particle of any one of embodiments 130-138, wherein the at least 7 consecutive amino acids comprise TQDWHRI (SEQ ID NO: 941).140. The AAV particle of any one of embodiments 130-134, wherein the at least 3 consecutive amino acids comprise TNT.141. The AAV particle of any one of embodiments 130-134 and 140, wherein the at least 4 consecutive amino acids comprise TNTQ (SEQ ID NO: 4688).142. The AAV particle of any one of embodiments 130-134, 140, and 141, wherein the at least 5 consecutive amino acids comprise TNTQD (SEQ ID NO: 5119).143. The AAV particle of any one of embodiments 130-134 and 140-142, wherein the at least 6 consecutive amino acids comprise TNTQDW (SEQ ID NO: 5120).144. The AAV particle of any one of embodiments 130-134 and 140-143, wherein the at least 7 consecutive amino acids comprise TNTQDWH (SEQ ID NO: 5121).145. The AAV particle of any one of embodiments 130-134 and 140-144, wherein the at least 8 consecutive amino acids comprise TNTQDWHR (SEQ ID NO: 4898).146. The AAV particle of any one of embodiments 130-134 and 140-145, wherein the at least 9 consecutive amino acids comprise TNTQDWHRI (SEQ ID NO: 746).147. The AAV particle of any one of embodiments 130-146, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two, or at least three but no more than four modifications relative to the amino acid sequence of any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291, 293, 301, 306, 307, 308, 309, 314, and 336.148. The AAV particle of any one of embodiments 130-147, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one. at least two. or at least three but no more than four modifications relative to the amino acid sequence of TQDWHRI (SEQ ID NO: 941).149. The AAV particle of any one of embodiments 130-147. wherein the AAV capsid variant comprises an amino acid sequence comprising at least one. at least two. or at least three but no more than four modifications relative to the amino acid sequence of TNTQDWHRI (SEQ ID NO: 746).150. The AAV particle of any one of embodiments 130-149, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two. or at least three but no more than four modifications relative to the amino acid sequence of ATNTQDWHRIAQT (SEQ ID NO: 744).151. The AAV particle of any one of embodiments 130-150, w herein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two, or at least three but no more than four substitutions relative to the amino acid sequence of any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291. 293, 301, 306, 307, 308, 309, 314, and 336.152. The AAV particle of any one of embodiments 130-149 and 151, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least tw o, or at least three but no more than four substitutions relative to the amino acid sequence of TQDWHRI (SEQ ID NO: 941).153. The AAV particle of any one of embodiments 130-149, 151, and 152, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two, or at least three but no more than four substitutions relative to the amino acid sequence of TNTQDWHRI (SEQ ID NO: 746).154. The AAV particle of any one of embodiments 130-153, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one. at least two. or at least three but no more than four substitutions relative to the amino acid sequence of ATNTQDWHRIAQT (SEQ ID NO: 744).155. The AAV particle of any one of embodiments 1-154, wherein the AAV capsid variant comprises the amino acid sequence of any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291, 293, 301, 306, 307, 308, 309, 314, and 336.156. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49. 51-69, 71-83, 85-99, and 101-155, wherein the AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941), optionally wherein the amino acid sequence replaces amino acids 584-590 of SEQ ID NO: 138.157. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83, 85-99, and 101-156. wherein the AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941), optionally wherein the amino acid sequence corresponds to amino acids 584-590 of SEQ ID NO: 981.158. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21. 23-39, 42-49, 51-69. 71-83, 85-99. 101-149, 151-153. and 155-157, wherein the AAV capsid variant comprises an amino acid sequence encoded by:(i) the nucleotide sequence of SEQ ID NO: 942;(ii) a nucleotide sequence comprising at least one, at least tw o. at least three, at least four, at least five, at least six, or at least seven modifications, but no more than ten modifications, relative to the nucleotide sequence of SEQ ID NO: 942; or(iii) a nucleotide sequence comprising at least one, at least two, at least three, at least four, at least five, at least six, or at least seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 942.159. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, 85-99, and 101-158, wherein the AAV capsid variant comprises an amino acid sequence encoded by:(i) the nucleotide sequence of SEQ ID NO: 747;(ii) a nucleotide sequence comprising at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, but no more than ten modifications, relative to the nucleotide sequence of SEQ ID NO: 747; or(iii) a nucleotide sequence comprising at least one. at least two. at least three, at least four, at least five, at least six, or at least seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 747.160. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, 85-99, and 101-149, 151-153, and 155-158, wherein the nucleotide sequence encoding the AAV capsid variant comprises:(i) the nucleotide sequence of SEQ ID NO: 942;(ii) a nucleotide sequence comprising at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, but no more than ten modifications, relative to the nucleotide sequence of SEQ ID NO: 942; or(iii) a nucleotide sequence comprising at least one. at least two. at least three, at least four, at least five, at least six, or at least seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 942.161. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83. 85-99, and 101-160. wherein the nucleotide sequence encoding the AAV capsid variant comprises:(i) the nucleotide sequence of SEQ ID NO: 747;(ii) a nucleotide sequence comprising at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, but no more than ten modifications, relative to the nucleotide sequence of SEQ ID NO: 747; or(iii) a nucleotide sequence comprising at least one, at least two. at least three, at least four, at least five, at least six, or at least seven, but no more than ten different nucleotides relative to the nucleotide sequence of SEQ ID NO: 747.162. The AAV particle of any one of embodiments 130-157, wherein the amino acid sequence provided in Table 1, 2A, 2B, 14, 27, 28, or 29 is present in loop VIII, optionally wherein loop VIII comprises amino acids 580-599, as numbered according to SEQ ID NO: 138 or 981.163. The AAV particle of any one of embodiments 130-157, wherein the amino acid sequence provided in Table 1, 2A, 2B, 14. 27, 28, or 29 replaces ammo acids 584, 585, 586, 587, 588, 589, and / or 590 (e.g., H584, Q585, S586, A587, Q588, A589, and / or Q590), as numbered according to the amino acid sequence of SEQ ID NO: 138.164. The AAV particle of any one of embodiments 130-157, wherein the amino acid sequence provided in Table 1, 2A, 2B, 14. 27, 28, or 29 is present at amino acids 584, 585. 586, 587, 588, 589, and / or 590, as numbered according to the amino acid sequence of SEQ ID NO: 981.165. The AAV particle of any one of embodiments 130-157. wherein the amino acid sequence provided in Table 1, 2A, 2B, 14. 27. 28, or 29 corresponds to amino acids 584-590 (e.g., T584, Q585. D586. W587, H588, R589. and / or 1590), numbered according to the amino acid sequence of SEQ ID NO: 981.166. The AAV particle of any one of embodiments 121-157, wherein the amino acid sequence provided in Table 1, 2A, 2B, 14. 27, 28, or 29 replaces amino acids 582, 583, 584, 585, 586, 587, 588. 589, and / or 590 (e.g., T582, N583, H584, Q585, S586, A587, Q588, A589, and / or Q590) of SEQ ID NO: 138.167. The AAV particle of any one of embodiments 121-157, wherein the amino acid sequence provided in Table 1, 2A, 2B, 14. 27, 28, or 29 is present at amino acids 582, 583. 584, 585, 586, 587, 588, 589, and / or 590, as numbered according to the amino acid sequence of SEQ ID NO: 981.168. The AAV particle of any one of embodiments 121-157. wherein the amino acid sequence provided in Table 1, 2A, 2B, 14. 27. 28, or 29 replaces amino acids 582. 583, 584, 585. 586, 587, 588. 589, 590. 591, 592, 593. 594, 595, and / or 596 (e.g.. T582. N583. H584, Q585, S586, A587. Q588. A589, Q590, A591. Q592, T593, G594, W595, and / or V5965) of SEQ ID NO: 138.169. The AAV particle of any one of embodiments 121-157, wherein the amino acid sequence provided in Table 1. 2A. 2B. 14, 27, 28, or 29 is present at amino acids 582, 583, 584. 585, 586. 587, 588, 589.590. 591, 592, 593. 594, 595, and / or 596, as numbered according to the amino acid sequence of SEQ ID NO: 981.170. The AAV particle of any one of embodiments 1-169, wherein the AAV capsid variant comprises one. two, three, four, five, or all of:(i) an amino acid other than H at position 584 (e.g., T);(ii) an amino acid other than S at position 586 (e.g., D);(iii) an amino acid other than A at position 587 (e.g., W);(iv) an amino acid other than Q at position 588 (e.g., H);(v) an amino acid other than A at position 589 (e.g., R); and / or(vi) an amino acid other than Q at position 590 (e.g., I); wherein the amino acids in (i)-(vi) are numbered according to SEQ ID NO: 138.171. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising one, two, three, four, five, or all of:(i) an amino acid other than H at position 584 (e.g., T);(ii) an amino acid other than S at position 586 (e.g., D);(iii) an amino acid other than A at position 587 (e.g.. W);(iv) an amino acid other than Q at position 588 (e.g.. H);(v) an amino acid other than A at position 589 (e.g., R); and / or(vi) an amino acid other than Q at position 590 (e.g., I); wherein the amino acids in (i)-(vi) are numbered according to SEQ ID NO: 138.172. The AAV particle of any one of embodiments 1-171, wherein the AAV capsid variant comprises one, two, three, four, five, or all of: T at position 584, D at position 586, W at position 587, H at position 588, R at position 589, and / or I at position 590, as numbered according to SEQ ID NO: 981.173. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising one, two, three, four, five, or all of: T at position 584, D at position 586. W at position 587, H at position 588, R at position 589. and / or I at position 590, as numbered according to SEQ ID NO: 981.174. The AAV particle of any one of embodiments 1-173, wherein the AAV capsid variant comprises one. two, three, four, five, or all of the substitutions H584T, S586D. A587W, Q588H, A589R, and / or Q590I, as numbered according to SEQ ID NO: 138.175. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g.. encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising one, two. three, four, five or all of the substitutions H584T, S586D, A587W, Q588H, A589R, and / or Q590I, as numbered according to SEQ ID NO: 138.176. The AAV particle of any one of embodiments 1-175, wherein the AAV capsid variant comprises an amino acid other than H at position 584 (e.g., T), an amino acid other than S at position 586 (e.g., D), an amino acid other than A at position 587 (e.g., W), an amino acid other than Q at position 588 (e.g., H), an amino acid other than A at position 589 (e.g., R), and an amino acid other than Q at position 590 (e.g., I), as numbered according to SEQ ID NO: 138.177. The AAV particle of any one of embodiments 1-176, wherein the AAV capsid variant comprises T at position 584, D at position 586, W at position 587, H at position 588, R at position 589, and I at position 590, as numbered according to SEQ ID NO: 981.178. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83, 85-98, and 100-164, wherein the AAV capsid variant comprises an amino acid at each of positions 584, 586, 587, 588. 589, and 590 that is different from the amino acid at the corresponding positions in SEQ ID NO: 138.179. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, 85-99, and 101-178, wherein the AAV capsid variant comprises an amino acid at each of positions 582, 583, 584, 586, 587, 588, and 589 that is different from the amino acid at the corresponding positions in SEQ ID NO: 138.180. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83, 85-99, and 101-179, wherein the AAV capsid variant comprises an amino acid at each of positions 582, 583, 584, 586. 587, 588, 589. 590, 591, 592, 593, 594. 595, and 596 that is different from the amino acid at the corresponding positions in SEQ ID NO: 138.181. The AAV particle of any one of embodiments 1-181, wherein the AAV capsid variant comprises the substitutions H584T. S586D. A587W, Q588H, A589R, and Q5901. as numbered according to SEQ ID NO: 138.182. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g.. encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising T at position 584. D at position 586, W at position 587, H at position 588, R at position 589, and I at position 590, as numbered according to SEQ ID NO: 981.183. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g.. encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising the substitutions H584T, S586D, A587W, Q588H, A589R, and Q590I, as numbered according to SEQ ID NO: 138.184. The AAV particle of any one of embodiments 1-183, wherein the AAV capsid variant further comprises an amino acid other than A position 581, as numbered according to SEQ ID NO: 138 or 981.185. The AAV particle of any one of embodiments 1-184, wherein the AAV capsid variant further comprises a T at position 581 or a V at position 581. as numbered according to SEQ ID NO: 138 or 981.186. The AAV particle of any one of embodiments 1-185, wherein the AAV capsid variant comprises the substitutions A581T or A581V, as numbered according to SEQ ID NO: 138 or 981.187. The AAV particle of any one of embodiments 1, 3-7, 9-15, 17-20, 22-38, 40-48, 50, 130-134, 147, 151, 155, 162. 163, 166-177. and 181-186, wherein the AAV capsid variant comprises an amino acid other than Q at position 585. as numbered according to SEQ ID NO: 138.188. The AAV particle of any one of embodiments 1, 3-7, 9-15, 17-20, 22-38, 40-48, 50, 130-134, 147, 151, 155, 162, 163, 166-177, and 181-187, wherein the AAV capsid variant comprises the amino acid K at position 585, as numbered according to SEQ ID NO: 138.189. The AAV particle of any one of embodiments 1-189, wherein the AAV capsid variant further comprises:(i) a modification in loop I, II, IV, and / or VI; and / or(ii) a substitution at position K449, e.g., a K449R substitution, as numbered according to SEQ ID NO: 138 or 981.190. The AAV particle of any one of embodiments 1-189, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two, or at least three modifications, but not more than 30. not more than 20, or not more than 10 modifications, relative to the amino acid sequence of SEQ ID NO: 138.191. The AAV particle of any one of embodiments 1-190, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two, or at least three, but no more than not more than 30, not more than 20, or not more than 10 substitutions relative to the amino acid sequence of SEQ ID NO: 138.192. The AAV particle of any one of embodiments 1-191, wherein the AAV capsid variant an amino acid sequence with at least 90% (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to SEQ ID NO: 138.193. The AAV particle of embodiment 192, wherein the AAV capsid variant an amino acid sequence with at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to SEQ ID NO: 138.194. The AAV particle of any one of embodiments 1-193, wherein the AAV capsid variant comprises an amino acid sequence encoded by a sequence with at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to SEQ ID NO: 137.195. The AAV particle of embodiment 194. wherein the AAV capsid variant comprises an amino acid sequence encoded by a sequence with at least 90% (e.g.. at least 90%, at least 91%, at least 92%. at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to SEQ ID NO: 137.196. The AAV particle of any one of embodiments 1-195, wherein the AAV capsid variant comprises a VP1 protein, a VP2 protein, and a VP3 protein in an about 1:1 :10 ratio.197. The AAV particle of any one of embodiments 1-196, wherein the AAV capsid variant comprises amino acids 138-736, e.g., a VP2, of SEQ ID NO: 981, or a sequence with at least 80% (e.g., at least 80%, at least 85%. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%. or at least 99%) sequence identity thereto.198. The AAV particle of any one of embodiments 1-197, wherein the AAV capsid variant comprises amino acids 203-736, e.g.. a VP3, of SEQ ID NO: 981, or a sequence with at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99%) sequence identity' thereto.199. The AAV particle of any one of embodiments 1-198, wherein the AAV capsid variant comprises an amino acid sequence that is at least 80% (e.g., at least 80%. at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99%) identical to amino acids 138-736, e.g.. a VP2, of SEQ ID NO: 138.200. The AAV particle of any one of embodiments 1-199, wherein the AAV capsid variant comprises an amino acid sequence that is at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to amino acids 203-736, e.g., a VP3, of SEQ ID NO: 138.201. The AAV particle of any one of embodiments 1-3, 5-8, 10-16, 18-21, 23-39, 42-49, 51-69, 71-83, 85-99, 101-186, and 189-200, wherein the AAV capsid variant comprises an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, or all 7 consecutive amino acids from the amino acid sequence of TQDWHRI (SEQ ID NO: 941), wherein:(i) the at least 3 consecutive amino acids comprise TQD:(ii) the at least 4 consecutive amino acids comprise TQDW (SEQ ID NO: 4684);(iii) the at least 5 consecutive amino acids comprise TQDWH (SEQ ID NO: 4685);(iv) the at least 6 consecutive amino acids comprise TQDWHR (SEQ ID NO: 4686); or(v) the 7 consecutive amino acids comprise TQDWHRI (SEQ ID NO: 941): wherein the AAV capsid variant comprises: (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 981; (b) a VP2 protein comprising amino acids 138-736 of SEQ ID NO: 981:(c) a VP3 protein comprising amino acids 203-736 of SEQ ID NO: 981; or (d) an amino acid sequence with at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to any one of tire amino acid sequences in (a)-(c).202. The AAV particle of embodiment 201, wherein the AAV capsid variant comprises an amino acid sequence with at least 90% (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to any one of the amino acid sequences in (a)-(c).203. The AAV particle of any one of embodiments 1-3, 5-8, 10-16. 18-21, 23-39, 42-49. 51-69, 71-83, 85-99, 101-186. and 189-202, wherein the AAV capsid variant comprises 1, 2, or 3 substitutions relative to the amino acid sequence of TQDWHRI (SEQ ID NO: 941). wherein the AAV capsid variant comprises an amino acid sequence at least 90% (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%. at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981.204. The AAV particle of any one of embodiments 1-203, wherein the AAV capsid variant comprises the amino acid sequence of any one of SEQ ID NO: 981, or an amino acid sequence with at least 90% (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto.205. The AAV particle of any one of embodiments 1-204, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two, or at least three modifications, but not more than 30, not more than 20 or not more than 10 modifications relative to the amino acid sequence of SEQ ID NO: 981.206. The AAV particle of any one of embodiments, 1-205, wherein the AAV capsid variant comprises an amino acid sequence comprising at least one, at least two or at least three, but not more than 30, not more than 20 or not more than 10 substitutions, relative to the amino acid sequence of SEQ ID NO: 981.207. The AAV particle of any one of embodiments 1-206, wherein the AAV capsid variant comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence with at least 80% (e.g.. at least 80%, at least 85%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99%) sequence identity thereto.208. The AAV particle of embodiment 207, wherein the AAV capsid variant comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983.209. The AAV particle of embodiment 207, wherein the AAV capsid variant is encoded by a codon- optimized nucleotide sequence.210. An AAV particle comprising an AAV capsid variant (e.g.. an AAV9 capsid variant) comprising the amino acid sequence of any one of embodiments 1-3. 5-8, 10-16, 18-21. 23-39, 42-49, 51-69. 71-83, 85- 99, 101-186. and 189, and further comprising an amino acid sequence at least 95% (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99%) identical to SEQ ID NO: 981.211. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g.. encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising the amino acid sequence of SEQ ID NO: 981.212. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.213. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising amino acids 203-736 of SEQ ID NO: 981, or an amino acid sequence at least 80% (e.g., at least 80%. at least 85%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99%) identical thereto, wherein the AAV capsid variant comprises T at position 584, D at position 586, W at position 587, H at position 588, R at position 589, and I at position 590, as numbered according to SEQ ID NO: 981.214. The AAV particle of embodiment 213, wherein the AAV capsid variant comprises amino acids 203- 736 of SEQ ID NO: 981.215. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g.. encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising amino acids 203-736 of SEQ ID NO: 981.216. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising amino acids 138-736 of SEQ ID NO: 981, or an amino acid sequence at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical thereto, wherein the AAV capsid variant comprises T at position 584, D at position 586, W at position 587, H at position 588, R at position 589, and I at position 590, as numbered according to SEQ ID NO: 981.217. The AAV particle any one of embodiments 213-216, wherein the AAV capsid variant comprises amino acids 138-736 of SEQ ID NO: 981.218. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g., encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising amino acids 138-736 of SEQ ID NO: 981.219. An AAV particle comprising a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence (e.g.. encoding a human GBA1 protein) and an AAV capsid variant (e.g., an AAV9 capsid variant) comprising the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence at least 80% (e.g., at least 80%, at least 85%, at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99%) identical thereto, wherein the AAV capsid variant comprises T at position 584, D at position 586, W at position 587, H at position 588, R at position 589, and I at position 590, as numbered according to SEQ ID NO: 981.220. The AAV particle any one of embodiments 213-219, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981.221. The AAV particle of any one of embodiments 1-220, wherein the AAV capsid variant has increased tropism for a CNS cell or tissue, e.g., a brain cell, brain tissue, spinal cord cell, or spinal cord tissue, relative to the tropism of an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138.222. The AAV particle of any one of embodiments 1-221, wherein the AAV capsid variant transduces a brain region, e.g., sensory cortex, motor cortex, putamen, thalamus, caudate, hippocampus, and / or cerebellum; optionally wherein the level of transduction is at least 39, at least 50, at least 100, at least 120, at least 132, at least 146, at least 150, at least 161, at least 174, at least 175, at least 200, at least 225, at least 250. at least 275. at least 283. at least 300. at least 350. at least 400. at least 450. at least 500. at least 525. at least 528. or at least 550-fold greater as compared to an AAV particle comprising an AAVcapsid comprising the amino acid sequence of SEQ ID NO: 138, e.g., when measured by an assay, e.g., an immuiiohistochcmistry assay or a qPCR or ddPCR assay, e.g., as described in Example 2.223. The AAV particle of any one of embodiments 1-222, which is enriched at least 10, at least 14, at least 20, at least 24, at least 50, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 425, at least 450, or at least 460-fold in the brain compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1 or 3.224. The AAV particle of any one of embodiments 1-223, which is enriched at least 200, at least 300, at least 400, at least 425, at least 450, or at least 460-fold in the brain compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138, e.g.. when measured by an assay as described in Example 1.225. The AAV particle of any one of embodiments 1-224, which is enriched in the brain of at least two or at least three species, e.g.. a non-human primate and rodent (e.g., mouse), e.g.. as compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138.226. The AAV particle of any one of embodiments 1-225, which is enriched at least 2. at least 3, at least 5, at least 10, at least 15, at least 20. at least 25, at least 30, at least 35, at least 40, at least 45. at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90, at least 95, at least 100, at least 105, at least 115, at least 120, at least 125, at least 130, at least 135, at least 140, at least 145, at least 150, at least 155, at least 160, at least 165, at least 170. at least 175. at least 180, at least 190, at least 200, at least 205, or at least 210-fold in the brain of at least tw o or at least three species, e.g., a non-human primate and rodent (e.g., mouse), compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1 or 4.227. The AAV particle of embodiment 225 or 226, wherein the at least two or at least three species comprise Macaca fascicularis, CMorocebus sabaeus, Callithrix jacchus, and / or mouse (e.g., outbred mice).228. The AAV particle of any one of embodiments 1-227, which is enriched at least 2, at least 3. at least 4. at least 5. at least 10, at least 15, at least 17. at least 20, at least 50, at least 75, at least 100, at least 103, at least 107. at least 125. at least 150. at least 200. at least 250. at least 300, at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1200-fold in the brain compared to an AAVparticle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 981, e.g., when measured by an assay as described in Example 3.229. The AAV particle of any one of embodiments 1-228, which delivers an increased level of GBA1 to a brain region, optionally wherein the GBA1 level is increased by at least 39, at least 50. at least 100. at least 120, at least 132, at least 146, at least 150, at least 161, at least 174, at least 175, at least 200, at least 225, at least 250, at least 275, at least 283, at least 300, at least 350, at least 400, at least 450, at least 500. at least 525, at least 528, or at least 550-fold as compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138, e.g., when measured by an assay, e.g., a qRT- PCR, a ddPCR, or a qPCR assay (e.g., as described in Example 2).230. The AAV particle of any one of embodiments 1-229, which delivers an increased level of viral genomes to a brain region, optionally wherein the level of viral genomes is increased by at least 2. at least 5, at least 7, at least 10, at least 15, at least 19, at least 20, at least 22, or at least 25-fold as compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138, e.g.. when measured by an assay, e.g.. a qRT-PCR or a qPCR assay (e.g., as described in Example 2).231. The AAV particle of embodiment 229 or 230. wherein the brain region is a sensory’ cortex, motor cortex, putamen, thalamus, caudate, hippocampus, and / or cerebellum.232. The AAV particle of any one of embodiments 1-231, which is enriched at least 5. at least 10, at least 50, at least 100, at least 115, at least 120, at least 150, at least 175, at least 200, at least 207, at least 225, at least 250, or at least 275-fold in the spinal cord compared to an AAV particle comprising an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138, e.g., when measured by an assay as described in Example 1 or 2.[Embodiments 233-311 are intentionally absent.]312. The AAV particle of any one of embodiments 1-232, wherein the viral genome is single-stranded.313. The AAV particle of any one of embodiments 1-232, wherein the viral genome is self- complementary.314. The AAV particle of any one of embodiments 1-232, embodiment 312, or embodiment 313, wherein the viral genome further comprises a nucleotide sequence encoding a Rep protein, e.g., a non-structural protein, wherein the Rep protein comprises a Rep78 protein, a Rep68 protein, a Rep52 protein, and / or a Rep40 protein (e.g., a Rep 78 protein and a Rep52 protein).315. The AAV particle of any one of embodiments 1-232, embodiment 312, or embodiment 313, wherein die AAV particle furdier comprises a nucleotide sequence encoding a Rep protein, e.g., a non-structural protein, wherein the Rep protein comprises a Rep78 protein, a Rep68 protein, a Rep52 protein, and / or a Rep40 protein (e.g., a Rep78 protein and a Rep52 protein).316. The AAV particle of embodiment 314 or 315, wherein the Rep78 protein, the Rep68 protein, the Rep52 protein, and / or the Rep40 protein is encoded by at least one Rep gene.317. The AAV particle of any one of embodiments 1-232 and 312-316. wherein the AAV particle is an isolated AAV particle.318. The AAV particle of any one of embodiments 1-232 and 312-317. wherein the AAV particle is a recombinant AAV particle.319. A cell, e.g., a host cell, comprising the AAV particle of any one of embodiments 1-232 and 312- 318.320. The cell of embodiment 319. wherein the cell is a mammalian cell or an insect cell.321. The cell of embodiment 319 or 320, wherein the cell is a cell of a brain region or a spinal cord region, optionally wherein the cell is a cell of the sensory cortex, motor cortex, putamen. thalamus, caudate, hippocampus, or cerebellum.[Embodiments 322-325 arc intentionally absent].326. A pharmaceutical composition comprising the AAV particle of any one of embodiments 1-232 and 312-318, and a pharmaceutically acceptable excipient.327. A method of delivering GBA1 to a cell or tissue (e.g.. a CNS cell or CNS tissue), comprising administering an effective amount of the pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318.328. The method of embodiment 327, wherein the cell is a cell of a brain region or a spinal cord region, optionally a cell of the frontal cortex, sensory cortex, motor cortex, caudate, cerebellar cortex, cerebral cortex, brain stem, hippocampus, or thalamus.329. The method of embodiment 327 or 328, wherein the cell is a neuron, a sensory neuron, and / or a motor neuron.330. The method of any one of embodiments 327-329, wherein the cell or tissue is within a subject.331. The method of embodiment 330, wherein the subject has, has been diagnosed with having, or is at risk of having a genetic disorder.332. The method of embodiment 330 or embodiment 331, wherein the subject has, has been diagnosed with having, or is at risk of having a neurological disorder.333. The method of embodiment 330 or embodiment 331, wherein the subject has, has been diagnosed with having, or is at risk of having a neurodegenerative disorder.334. The method of embodiment 330 or embodiment 331, wherein the subject has. has been diagnosed with having, or is at risk of having a muscular disorder or a neuromuscular disorder.335. A method of treating a subject having or diagnosed with having a genetic disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318.336. A method of treating a subject having or diagnosed with having a neurological disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318.337. A method of treating a subject having or diagnosed with having a muscular disorder or a neuromuscular disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318.338. A method of treating a subject having or diagnosed with having a neurodegenerative disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318.339. The method of any one of embodiments 327-338. wherein the genetic disorder, neurological disorder, neurode generative disorder, muscular disorder, or neuromuscular disorder is Parkinson’s Disease (PD) (e.g., a PD associated with a GBA1 mutation). Parkinson’s Disease Dementia (PDD),Gaucher Disease (GD) (e.g., GDI, GD2, or GD3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Spinal muscular atrophy (SMA), Multiple System Atrophy (MSA), or Multiple sclerosis (MS).340. The method of any one of embodiments 335-339, where treating comprises prevention of progression of the disorder in the subject.341. The method of any one of embodiments 330-340. wherein the subject is a human.342. The method of any one of embodiments 330-341. wherein the AAV particle or the pharmaceutical composition is administered to the subject intravenously, via intra-cisterna magna injection (ICM). intracerebrally, intrathecally. intracerebroventricularly, via intraparenchymal administration, intraarterially, or intramuscularly.343. The method of any one of embodiments 330-342, wherein the AAV particle or pharmaceutical composition is administered to the subject via focused ultrasound (FUS), e g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration.344. The method of any one of embodiments 330-343, wherein the AAV particle or pharmaceutical composition is administered to the subject intravenously.345. The method of any one of embodiments 330-344, wherein the AAV particle or pharmaceutical composition is administered to the subject via intra-cistema magna injection (ICM).346. The method of any one of embodiments 330-345, wherein the AAV particle or pharmaceutical composition is administered to the subject intraarterially.347. The method of any one of embodiments 342-346, wherein administration of the AAV particle or pharmaceutical composition results in an increased expression of a GBA1 gene. mRNA. protein, or combination thereof.348. The method of any one of embodiments 342-346. wherein administration of the AAV particle or pharmaceutical composition results in an increased activity of a GBA1 gene, mRNA, protein, or a combination thereof.349. The pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318, for use in a method of delivering GBA1 to a cell or tissue.350. The pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318, for use in a method of treating a genetic disorder, a neurological disorder, a neurodegenerative disorder, a muscular disorder, or a neuromuscular disorder.351. The pharmacal composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318, for use in the manufacture of a medicament.352. Use of the pharmacal composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318 in the manufacture of a medicament.353. Use of the pharmaceutical composition of embodiment 326 or the AAV particle of any one of embodiments 1-232 and 312-318 in the manufacture of a medicament for treating a genetic disorder, a neurological disorder, a neurodegenerative disorder, a muscular disorder, or a neuromuscular disorder.354. An adeno-associated virus (AAV) particle comprising an AAV capsid variant and a viral genome, wherein the viral genome comprises a P-glucocerebrosidase 1 (GBAl)-encoding sequence and the AAV capsid variant comprises an amino acid sequence having the formula [N1]-[N2]-[N3] (SEQ ID NO: 4681) in loop VIII: wherein [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682); and wherein:(i) [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4is Q, K, E, S, P, R, N, or H; and / or(ii) [N3] comprises amino acids X5, Xg, and X-, wherein X, is I, V, T, M, S, N, L, or F.355. The AAV particle of embodiment 354, wherein the AAV capsid variant is an AAV9 capsid variant.356. The AAV particle of embodiment 354 or embodiment 355, wherein the AAV capsid variant comprises an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to amino acids 203-736 of SEQ ID NO: 981.357. The AAV particle of any one of embodiments 354-356. wherein loop VIII comprises amino acids 580-599 as numbered according to SEQ ID NO: 981.358. The AAV particle of any one of embodiments 354-357. wherein:(i) Xi is T, S, R, A, I, C, N, K, L, or Q;(ii) X2is N, T, G, V, S, Y, K, I, H, D, or F; and(iii) X3is T, N, K, D, I. S, P, A, Y, E, V, L, M, R, H, Q, or C.359. The AAV particle of any one of embodiments 354-358, wherein:(i) X6is A, Y, P, N, S, T, G, E, V, W, F, or Q; and(ii) X7is Q, G. N, K, H, R, E. L, P, or M.360. The AAV particle of any one of embodiments 354-359. wherein [Nl] comprises the amino acid sequence of TNTQ (SEQ ID NO: 4688).361. The AAV particle of any one of embodiments 354-360. wherein LN3 J comprises the amino acid sequence of IAQ.362. The AAV particle of any one of embodiments 354-361, wherein [N1]-N2]-[N3] comprises the amino acid sequence of TNTQDWHRI AQ (SEQ ID NO: 343).363. The AAV particle of any one of embodiments 354-362, wherein:(i) [Nl] is present at amino acids 582-585;(ii) [N2] is present at amino acids 586-589; and(iii) [N3] is present at amino acids 590-592; wherein the amino acids are numbered according to SEQ ID NO: 981.364. The AAV particle of any one of embodiments 354-363, further comprising [N4] comprising amino acids X8, X9, Xio, and Xu, wherein:(i) X8is T, S, N, P, A, or I;(ii) X9is G, N, D, R, V, A, S, or Q;(iii) Xiois W, S, C. R, L, or G; and / or(iv) Xu is V, A, S, I, C, G, D, F, L, or T.365. The AAV particle of embodiment 364. wherein [N4] comprises the amino acid sequence of TGWV (SEQ ID NO: 5066).366. The AAV particle of embodiment 364 or embodiment 365, wherein [N4] is present at amino acids 593-596 as numbered according to SEQ ID NO: 981.367. The AAV particle of any one of embodiments 354-366, wherein [N1]-[N2]-[N3]-[N4] comprises the amino acid sequence of TNTQDWHRIAQTGWV (SEQ ID NO: 201).368. An adeno-associated virus (AAV) particle comprising:(i) a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence: and(ii) an AAV capsid variant comprising an amino acid sequence at least 95% (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99%) identical to amino acids 203-736 of SEQ ID NO: 981, wherein the AAV capsid variant comprises:T at position 584,D at position 586,W position 587,H at position 588,R at position 589, andI at position 590; wherein the amino acids are numbered according to SEQ ID NO: 981.369. The AAV particle of any one of embodiments 354-368, wherein the AAV capsid variant comprises an amino acid sequence having at least 95% (e g., at least 95%, at least 96%, at least 97%, at least 98%. or at least 99%) identity to amino acids 138-736 of SEQ ID NO: 981.370. An adeno-associated virus (AAV) particle comprising:(i) a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence; and(ii) an AAV capsid variant comprising an amino acid sequence at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to amino acids 138-736 of SEQ ID NO: 981, wherein the AAV capsid variant comprises:T at position 584,D at position 586,W position 587,H at position 588,R at position 589, andI at position 590; wherein the amino acids are numbered according to SEQ ID NO: 981.371. The AAV particle of any one of embodiments 354-370. wherein the AAV capsid variant comprises an amino acid sequence having at least 95% (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99%) identity to SEQ ID NO: 981.372. An adeno-associated virus (AAV) particle comprising:(i) a viral genome comprising a -glucocerebrosidase 1 (GBAl)-encoding sequence; and(ii) an AAV capsid variant comprising an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to SEQ ID NO: 981, wherein the AAV capsid variant comprises:T at position 584,D at position 586,W position 587,H at position 588,R at position 589, andI at position 590; wherein the amino acids are numbered according to SEQ ID NO: 981.373. The AAV particle of embodiment 372, wherein the AAV capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 99% identity to SEQ ID NO: 981;(ii) a VP2 protein comprising an amino acid sequence having at least 99% identity to amino acids 138-736 of SEQ ID NO: 981; and / or(iii) a VP3 protein comprising an amino acid sequence having at least 99% identity to amino acids 203-736 of SEQ ID NO: 981.374. The AAV particle of any one of embodiments 368-373, wherein the AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941).375. The AAV particle of any one of embodiments 368-374, wherein the AAV capsid variant comprises at least 8, at least 9, or at least 10 consecutive amino acids from the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343).376. The AAV particle of any one of embodiments 368-375, wherein the AAV capsid variant comprises the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343) at amino acids 582-592 as numbered according to SEQ ID NO: 981.377. The AAV particle of any one of embodiments 374-376, wherein the amino acid sequence of TQDWHRI (SEQ ID NO: 941) or TNTQDWHRIAQ (SEQ ID NO: 343) is present in loop VIII, wherein loop VIII comprises amino acids 580-599 as numbered according to SEQ ID NO: 981.378. The AAV particle of any one of embodiments 354-377. wherein the AAV capsid variant comprises:(i) a VP1 protein comprising or consisting of the amino acid sequence of SEQ ID NO: 981;(ii) a VP2 protein comprising or consisting of the amino acid sequence according to positions 138-736 of SEQ ID NO: 981; and / or(iii) a VP3 protein comprising or consisting of the amino acid sequence according to positions 203-736 of SEQ ID NO: 981.379. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1775 or an amino acid sequence at least 90% identical thereto.380. The AAV particle of any one of embodiments 1-232, 312-318, and 354-379, wherein the viral genome encodes a wildtvpc GBA1 protein.381. The AAV particle of any one of embodiments 1-232, 312-318, and 354-380, wherein the viral genome encodes a human GBA1 protein, a dog GBA1 protein, or an equine GBA1 protein.382. The AAV particle of any one of embodiments 1-232, 312-318. and 354-381, wherein the viral genome encodes a wildty pe human GBA1 protein.383. The AAV particle of any one of embodiments 1-232, 312-318. and 354-382, wherein the viral genome does not encode a hemagglutinin (HA) tag.384. The AAV particle of any one of embodiments 1-232, 312-318, and 354-383, wherein the GBA1- encoding sequence comprises SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.385. The AAV particle of any one of embodiments 1-232, 312-318, and 354-384, where the viral genome further comprises a signal sequence comprising the nucleotide sequence of SEQ ID NO: 2005 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.386. The AAV particle of any one of embodiments 1-232, 312-318, and 354-385, wherein the viral genome comprises the nucleotide sequence of 2001 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto.387. The AAV particle of any one of embodiments 1-232, 312-318, and 354-386, wherein the viral genome encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1774 or an amino acid sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.388. The AAV particle of any one of embodiments 1-232, 312-318, and 354-387, wherein the viral genome comprises a promoter operably linked to the GBAl-encoding sequence.389. The AAV particle of embodiment 388. wherein the promoter comprises an EF-la promoter, a CBA promoter, a chicken |3-actin (CBA) promoter, a CAG promoter, a CAG derivative promoter, a CMV immediate-early enhancer and / or promoter, a CMV promoter, a P glucuronidase (GUSB) promoter, a ubiquitin C (UBC) promoter, a neuron-specific enolase (NSE) promoter, a platelet-derived growth factor (PDGF) promoter, a platelet-derived growth factor B-chain (PDGF- ) promoter, an intercellular adhesion molecule 2 (ICAM-2) promoter, a synapsin (Syn) promoter, a methyl-CpG binding protein 2 (MeCP2) promoter, a Ca2+ / calmodulin-dependent protein kinase II (CaMKII) promoter, a metabotropic glutamate receptor 2 (mGluR2) promoter, a neurofilament light (NFL) or heavy (NFH) promoter, a -globin minigene n 2 promoter, a preproenkephalin (PPE) promoter, an enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2) promoter, a glial fibri 1 lan acidic protein (GFAP) promoter, a myelin basic protein (MBP) promoter, a cardiovascular promoter (e.g., aMHC. cTnT, and CMV-MLC2k). a liver promoter (e.g.. hAAT, TBG). a skeletal muscle promoter (e.g., desmin, MCK. C512), or a fragment, e.g., a truncation, or a functional variant thereof.390. The AAV particle of embodiment 388 or embodiment 389, wherein the promoter comprises the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto.391. The AAV particle of any one of embodiments 1-232, 312-318, and 354-390, wherein the viral genome further comprises an enhancer.392. The AAV particle of embodiment 391. wherein the enhancer comprises a CMV immediate-early (CMVie) enhancer; optionally wherein the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.393. The AAV particle of any one of embodiments 1-232, 312-318, and 354-392, wherein the viral genome further comprises an intron; optionally wherein the intron comprises die nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.394. The AAV particle of any one of embodiments 1-232, 312-318, and 354-393, wherein the viral genome further comprises a polyadenylation (poly A) region; optionally wherein the polyA region comprises the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto.395. The AAV particle of any one of embodiments 1-232. 312-318, and 354-394. wherein the viral genome further comprises an inverted terminal repeat (ITR); optionally wherein the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830.396. The AAV particle of embodiment 395, wherein the viral genome comprises a 5’ ITR and a 3’ ITR. wherein the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto, and the 3‘ ITR comprises the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto.397. The AAV particle of any one of embodiments 1-232, 312-318, and 354-396, wherein the viral genome further comprises one or more miR183 binding sites.398. The AAV particle of any one of embodiments 1-232, 312-318, and 354-397, wherein the viral genome comprises at least four miR183 binding sites.399. The AAV particle of embodiment 398. wherein each of the at least four miR183 binding sites is separated by a spacer.400. The AAV particle of any one of embodiments 396-399, wherein each of the miR183 binding sites comprises the nucleotide sequence of SEQ ID NO: 1847 or a nucleotide sequence that has up to three modifications relative to SEQ ID NO: 1847.401. The AAV particle of any one of embodiments 1-232, 312-318, and 354-396, wherein the viral genome further comprises a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto.402. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) a GBA1 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; and(iv) a 3’ ITR.403. The AAV particle of any one of embodiments 1-232, 312-318. and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(v) a 3’ ITR.404. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) an intron;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vi) a 3‘ ITR.405. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CM Vie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vi) a 3’ ITR.406. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a poly adenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vii) a 3’ ITR.407. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5' inverted tenninal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto;(ii) a CMV immediate-early (CM Vie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.408. The AAV particle of embodiment 407. wherein:(i) the 5' ITR comprises the nucleotide sequence of SEQ ID NO: 1829;(ii) the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831;(iii) the CBA promoter comprises the nucleotide sequence of SEQ ID NO: 1834;(iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842;(v) the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001;(vi) the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846; and (viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.409. The AAV particle of any one of embodiments 1-232, 312-318, and 354-407, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.410. The AAV particle of embodiment 409. wherein the viral genome comprises or consists of the nucleotide sequence of SEQ ID NO: 2006.411. The AAV particle of any one of embodiments 1-232, 312-318. and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) at least one miR183 binding site; and(v) a 3’ ITR.412. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(iv) at least one miR183 binding site series comprising at least one miR183 binding site and at least one spacer sequence; and(v) a 3’ ITR.413. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(v) at least one miR183 binding site series comprising at least one miR183 binding site and at least one spacer sequence; and(vi) a 3’ lTR.414. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) an intron;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) at least one miR183 binding site series comprising at least one miR183 binding site and at least one spacer sequence; and(vii) a 3’ ITR.415. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto:(vi) at least one miR183 binding site; and(vii) a 3’ ITR.416. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a CMV immediate-early (CM Vie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vii) a 3’ ITR.417. The AAV particle of any one of embodiments 1-232, 312-318, and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%. at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(vi) at least one miR183 binding site comprising the nucleotide sequence of SEQ ID NO: 1847;(vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.418. The AAV particle of any one of embodiments 1-232, 312-318. and 354-378, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(ii) a CMV immediate-early (CMVie) enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a chicken beta actin (CBA) promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) a GBAl-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence at least 90% identical (e.g.. at least 90%, at least 91%. atleast 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a miR183 binding site series comprising the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.419. The AAV particle of embodiment 418. wherein:(i) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829;(ii) the CMVie enhancer comprises the nucleotide sequence of SEQ ID NO: 1831;(iii) the CBA promoter comprises the nucleotide sequence of SEQ ID NO: 1834;(iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842;(v) the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001;(vi) the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849;(vii) the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846; and(viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.420. The AAV particle of any one of embodiments 1-232, 312-318, and 354-418, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.421. The AAV particle of embodiment 420, wherein the viral genome comprises or consists of the nucleotide sequence of SEQ ID NO: 2007.422. A cell comprising the AAV particle of any one of embodiments 1-232. 312-318, and 354-421, optionally wherein the cell is a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an SI9 cell), or a bacterial cell.423. A method of making the AAV particle of any one of embodiments 1-232, 312-318. and 354-421, the method comprising:(i) providing a cell comprising a viral genome comprising a GBA1 -encoding sequence and a nucleic acid encoding an AAV capsid variant; and(ii) incubating the cell under conditions suitable to encapsulate the viral genome in the AAV capsid variant; thereby making the AAV particle.424. The method of embodiment 423. wherein the viral genome comprises:(a) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; or(b) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981.425. The method of embodiment 423 or embodiment 424, further comprising, prior to step (i), introducing a nucleic acid comprising the viral genome into the cell.426. The method of any one of embodiments 423-425, further comprising, prior to step (i). introducing the nucleic acid encoding the AAV capsid variant into the cell.427. The method of any one of embodiments 423-426, wherein the cell comprises a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an S19 cell), or a bacterial cell.428. A pharmaceutical composition comprising the AAV particle of any one of embodiments 1-232, 312- 318, and 354-421 and a pharmaceutically acceptable excipient.429. A method of delivering a GBA1 protein to a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318, and 354-421, thereby delivering the GBA1 protein.430. The method of embodiment 429, wherein the subject has, has been diagnosed with having, or is at risk of having a GBAl-related disorder.431. A method of treating a GBAl-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232. 312-318, and 354-421. thereby treating the GBAl-related disorder.432. The method of embodiment 431, wherein the subject has, has been diagnosed with having, or is at risk of having a GBA1 -related disorder.433. The method of any one of embodiments 430-432, wherein the GBA1 -related disorder is a GBA1- related neurodegenerative or neuromuscular disorder.434. The method of any one of embodiments 430-433. wherein the GBAl-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g.. GD type 1, 2, or 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD). Multiple System Atrophy (MSA), Alzheimer’s Disease (AD). Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome.435. The method of any one of embodiments 430-434, wherein the GBAl-related disorder is PD.436. The method of any one of embodiments 430-434, wherein the GBAl-related disorder is LBD.437. The method of any one of embodiments 430-434, wherein the GBAl-related disorder is GD (e g.,GD type 1 (GDI) or GD type 3 (GD3)).438. The method of any one of embodiments 430-434, wherein the GBAl-related disorder is DLB.439. A method of treating Parkinson’s Disease (PD) in a subject, comprising administering to the subject an effective amount of die pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318, and 354-421, thereby treating the PD.440. The method of embodiment 439, wherein the subject has, has been diagnosed with having, or is at risk of having PD.441. A method of treating Lewy Body Dementia (LBD) in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318. and 354-421, thereby treating the LBD.442. The method of embodiment 441, wherein the subject has, has been diagnosed with having, or is at risk of having LBD.443. A method of treating Gaucher Disease (GD) in a subject, comprising administering to tire subject an effective amount of the pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318, and 354-421, thereby treating the GD.444. The method of embodiment 443, wherein the subject has, has been diagnosed with having, or is at risk of having GD (e.g., GDI or GD3).445. A method of treating Dementia with Lewy Bodies (DLB) in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318. and 354-421, thereby treating the DLB.446. The method of embodiment 445, wherein the subject has. has been diagnosed with having, or is at risk of having DLB.447. The method of any one of embodiments 429-446, wherein the subject has one or more mutations in the GBA1 gene.448. The method of any one of embodiments 429-447, wherein, prior to the administration, the subject has lower GCase activity’ as compared to GCase activity in an individual who does not have a GBA1- related disorder, optionally wherein the level of GCase activity’ is measured by a 4-MUG assay or a SensoLyte Blue Glucocerebrosidase assay.449. The method of any one of embodiments 431-438, wherein the treating results in prevention of progression of the disorder in the subject.450. The method of any one of embodiments 431-438 or embodiment 449, wherein the treating results in amelioration of at least one symptom of the disorder in the subject.451. The method of embodiment 450, wherein the at least one symptom comprises developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, generalized weakness, trembling of a limb, depression, visual hallucinations, cognitive decline, or a combination thereof.452. The method of embodiment 449 or embodiment 450, wherein the treating results in a change in one or more biomarkers comprising a GCase activity, a level of glucocerebroside and other glycolipids, (e.g.. within immune cells such as macrophages), a level of synuclein aggregates (e.g., Lewy bodies), or a combination thereof.453. The method of any one of embodiments 429-452, wherein the subject is a human subject.454. The method of any one of embodiments 429-453, wherein the AAV particle or the pharmaceutical composition is delivered to a cell or tissue of the CNS.455. The method of embodiment 454, wherein the cell or tissue of the CNS is a cell or tissue of the amygdala, brainstem, caudate, central grey, cerebellum (e.g.. Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus. oculomotor nucleus, putamen. substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region).456. The method of any one of embodiments 429-453, wherein the AAV particle or the pharmaceutical composition is delivered to a peripheral cell or tissue.457. The method of embodiment 456, wherein the peripheral cell or tissue is a cell or tissue of the liver, heart, skeletal muscle, sympathetic ganglia, and / or plasma.458. The method of any one of embodiments 429-457, further comprising evaluating, e.g., measuring, the level of GBA1 expression (e.g.. GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) in the subject, e.g., in a cell, tissue, or fluid of the subject.459. The method of embodiment 458, wherein the level of GBA1 protein expression is measured by an ELISA, a Western blot, or an immunohistochemistry assay.460. The method of embodiment 458 or embodiment 459, wherein evaluating the subject’s level of GBA1 expression is performed prior to and / or subsequent to administration of the pharmaceutical composition or AAV particle, optionally wherein the subject’s level of GBA1 expression prior to administration is compared to the subject’s level of GBA1 expression subsequent to administration.461. The method of any one of embodiments 458-460. comprising evaluating the level of GBA1 expression in a cell or tissue of the central nervous system.462. The method of any one of embodiments 458-461, wherein the subject’s level of GBA1 protein expression subsequent to administration is increased relative to the subject’s level of GBA1 protein expression prior to administration.463. The method of any one of embodiments 429-462, further comprising evaluating, e.g., measuring, the level of GCase activity in the subject.464. The method of any one of embodiments 429-463. wherein the administration results in an increase in:(i) GCase activity in a cell, tissue, (e.g., a cell or tissue of the CNS, e.g.. amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g.. frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)), and / or fluid (e g., CSF and / or serum) of the subject relative to baseline and / or relative to GCase activity in a cell, tissue, or fluid of an individual with a GBA1 -related disorder who has not been administered the pharmaceutical composition or AAV particle;(ii) the number and / or level of viral genomes (VG) per cell in a CNS tissue (e.g.. amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gy rus, hippocampus, inferior colliculus, inferior olivary' complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)) of the subject relative to the number and / or level of VG per cell in a peripheral tissue of the subject; and / or(iii) GBA1 mRNA expression in a cell or tissue (e g., a cell or tissue of the CNS, e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus. oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g.. cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)) of the subject relative to baseline and / or relative to GBA1 mRNA expression in a cell or tissue of an individual with a GBA1 -related disorder who has not been administered the pharmaceutical composition or AAV particle.465. The method of any one of embodiments 429-464, further comprising administering to the subject at least one additional therapeutic agent and / or therapy.466. The method of embodiment 465. wherein the at least one additional therapeutic agent and / or therapy comprises an agent and / or therapy for treating a GBA1 -related disorder.467. The method of embodiment 466. wherein the at least one additional therapeutic agent and / or therapy for treating the GBAl-related disorder comprises enzyme replacement therapy (ERT) (e.g., imiglucerase. velaglucerase alfa. or taliglucerase alfa); substrate reduction therapy (SRT) (e.g., eliglustat or miglustat), levodopa, carbidopa. Safinamide. a dopamine agonist (e.g., pramipexole, rotigotine. or ropinirole). a dopamine antagonist (e.g., quetiapine, clozapine), an anticholinergic (e.g., benztropine or trihexyphenidyl), a cholinesterase inhibitor (e.g., rivastigmine, donepezil. or galantamine), an N-methyl- d-aspartate (NMDA) receptor antagonist (e.g.. memantine), or a combination thereof.468. The method of embodiment 466 or embodiment 467, wherein the at least one additional therapeutic agent and / or therapy is for treating Parkinson’s Disease (PD). Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1. 2. or 3). Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Nemodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome.469. The method of embodiment 468, wherein the GBAl-related disorder is PD, LBD, GD, or DLB, wherein, optionally, the GD is GD type 1 (GDI) or GD ty pe 3 (GD3).470. The method of any one of embodiments 429-469, further comprising administering an immunosuppressant to the subject.471. The method of embodiment 470. wherein the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, my cophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxychloroquine.472. The method of any one of embodiments 429-471. further comprising administering a blood transfusion to the subject.473. The pharmacal composition of embodiment 428 or the AAV particle of any one of embodiments 1-232. 312-318, and 354-421 for use in a method of treating a disorder according to any one of embodiments 431-472.474. The pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318, and 354-421 for use in the treatment of a GBAl-related disorder in a subject.475. The pharmacal composition or AAV particle for use of embodiment 474, wherein the GBAl- related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, 2. or 3). Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD). Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure. Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden-Spatz Syndrome.476. The pharmaceutical composition or AAV particle for use of embodiment 474 or embodiment 475. wherein the GBAl-related disorder is PD.477. The pharmaceutical composition or AAV particle for use of embodiment 476. wherein the subject has, has been diagnosed with having, or is at risk of having PD.478. The pharmaceutical composition or AAV particle for use of embodiment 474 or embodiment 475, wherein the GBAl-related disorder is LBD.479. The pharmaceutical composition or AAV particle for use of embodiment 478, wherein the subject has, has been diagnosed with having, or is at risk of having LBD.480. The pharmaceutical composition or AAV particle for use of embodiment 474 or embodiment 475, wherein the GBAl-related disorder is GD (e.g., GDI, GD2, or GD3).481. The pharmacal composition or AAV particle for use of embodiment 480, wherein the subject has. has been diagnosed with having, or is at risk of having GD (e.g., GDI or GD3).482. The pharmaceutical composition or AAV particle for use of embodiment 474 or embodiment 475. wherein the GBAl-related disorder is DLB.483. The pharmacal composition or AAV particle for use of embodiment 482, wherein the subject has. has been diagnosed with having, or is at risk of having DLB.484. Use of the pharmaceutical composition of embodiment 428 or the AAV particle of any one of embodiments 1-232, 312-318, and 354-421 in the manufacture of a medicament for the treatment of a GBA1 -related disorder in a subject.485. The use of embodiment 484, wherein the GBA1 -related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g., GD type 1, 2, or 3), Dementia with Lewy Bodies (DLB), Le Body Dementia (LBD), Multiple System Atrophy (MSA). Alzheimer’s Disease (AD). Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden-Spatz Syndrome.486. The use of embodiment 484 or embodiment 485, wherein the GBAl-related disorder is PD.487. The use of embodiment 486, wherein the subject has. has been diagnosed with having, or is at risk of having PD.488. The use of embodiment 484 or embodiment 485, wherein the GBAl-related disorder is GD (e.g., GDI or GD3).489. The use of embodiment 488. wherein the subject has, has been diagnosed with having, or is at risk of having GD (e.g., GDI or GD3).490. The use of embodiment 484 or embodiment 485, wherein the GBAl-related disorder is LBD.491. The use of embodiment 490. wherein the subject has, has been diagnosed with having, or is at risk of having LBD.492. The use of embodiment 484 or embodiment 485, wherein the GBAl-related disorder is DLB.493. The use of embodiment 492, wherein the subject has, has been diagnosed with having, or is at risk of having DLB.

[0074] The details of various aspects or embodiments of the present disclosure are set forth below. Other features, objects, and advantages of the disclosure will be apparent from the description and the claims. In the description, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art in the field of this disclosure. In the case of conflict, the present description will control.BRIEF DESCRIPTION OF THE DRAWINGS

[0075] FIG. 1 depicts the biodistribution (VG / cell) hi the cortex, striatum, and brainstem of wildtype mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812), VOY101.GBA VG35 (SEQ ID NO: 2006) or VOY101.GBA VG36 (SEQ ID NO: 2007).

[0076] FIG. 2A and FIG. 2B depict the GCase activity in the cortex, striatum, and brainstem of wild-type mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812), VOY101.GBA VG35 (SEQ ID NO: 2006) or VOY101.GBA VG36 (SEQ ID NO: 2007).

[0077] FIG. 3A and FIG. 3B depict the biodistribution, mRNA expression, and Gcase activities in the brainstem and DRGs of wild-type mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812), VOY101.GBA_VG35 (SEQ ID NO: 2006) or VOY101.GBA_VG36 (SEQ ID NO: 2007).

[0078] FIG. 4A and FIG. 4B depict the substrate quantification of glucosylceramide and glucosylsphingosine by LC-MS / MS in the brainstem, striatum, and DRGs of wild-ty pe mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812), VOY101.GBA VG35 (SEQ ID NO: 2006) or VOY101.GBA VG36 (SEQ ID NO: 2007).

[0079] FIG. 5A and FIG. 5B depict biodistribution (VG / cell) in the cortex and GCase activities in the cortex, striatum, and brainstem of wild-type mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812) or VOY101.GBA VG17-HA.

[0080] FIG. 6A depicts immunohistochemical analysis of HA expression in the cortex, striatum, and brainstem of wild-type mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812) or VOY101.GBA VG17-HA. FIG. 6B depicts immunohistochemical analysis of HA expression in the cerebellum, thalamus, and hippocampus of wild-type mice at 28 days post-IV injection of VOY101.GBA VG17 (SEQ ID NO: 1812) or VOY101.GBA VG17-HA.DETAILED DESCRIPTIONOverview

[0081] Described herein, inter alia, are compositions comprising an AAV capsid variant for deliver}' of a sequence encoding GBA1 protein, e.g., a wildtype GBA1 protein, e.g., a wildtype human GBA1 protein. In some embodiments, the present disclosure provides a method for delivering the AAV capsid variant comprising the sequence encoding the GBA1 protein to a cell or tissue in a subject. In some embodiments, the present disclosure provides a method for delivering the AAV capsid variant, thereby providing a GBA1 protein, e.g., wildtype GBA1 protein, e.g., a wildty pe human GBA1 protein, to a cell or tissue in a subject. The AAV capsid variants described herein have enhanced CNS tropism compared to other cells or tissues in the body, e g., liver and / or the DRG.

[0082] AAVs have proven to be useful as a biological tool due to their relatively simple structure, their ability to infect a wide range of cells (including quiescent and dividing cells) without integration into the host genome and without replicating, and their relatively benign immunogenic profile. Engineered adeno-associated virus (AAV) capsids with improved brain tropism represent an attractivesolution to the limitations of CNS delivery'. AAV-derived vectors are promising tools for clinical gene transfer because of their non-pathogenic nature, their low immunogenic profile, low rate of integration into the host genome, and long-term transgene expression in non-dividing cells. However, the transduction efficiency of naturally occurring AAVs in certain organs is too low for clinical applications, and capsid neutralization by pre-existing neutralizing antibodies may prevent treatment of a large proportion of patients. For these reasons, considerable efforts have been devoted to obtaining capsid variants with enhanced properties. Of many approaches tested so far, significant advances have resulted from directed evolution of AAV capsids using in vitro or in vivo selection of capsid variants created by capsid sequence randomization using either error-prone PCR, shuffling of various parent serotypes, or insertion of fully randomized short peptides at defined positions.

[0083] The genome of the virus may be modified to contain a minimum of components for the assembly of a functional recombinant virus, or viral particle, which is loaded with or engineered to target a particular tissue and express or deliver a desired payload. The genome of the virus may encode a GBA1 protein, and the viral particle comprising said genome may be delivered to a target cell, tissue, or organism. In some embodiments, the genome encodes a human GBA1 protein, e.g., a wildtype human GBA1 protein. In some embodiments, the target cell is a CNS cell. In some embodiments, the target tissue is a CNS tissue. In some embodiments, the target CNS tissue is brain tissue.

[0084] In some embodiments, the genome encodes a wildtype GBA1 protein. In some embodiments, the genome comprises a codon-optimized. CpG-reduced (e.g., CpG-depleted) nucleotide sequence encoding a wildtype GBA1 protein, e.g., as compared to a wildtype nucleotide sequence encoding a wildtype GBA1 protein (e.g., comprising the nucleotide sequence of SEQ ID NO: 1776 or 1777). In some embodiments, the target cell is a CNS cell. In some embodiments, the target tissue is a CNS tissue. The target CNS tissue may be brain tissue. In some embodiments, the CNS tissue comprises amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory' cortex, temporal cortex, visual cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary' complex, nucleus ambiguus. oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g.. cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region).

[0085] Gene therapy presents an alternative approach for treating a GBAl-related disorder, e.g., Parkinson’s Disease (PD) or Gaucher Disease (GD). AAVs are commonly used in gene therapy approaches as a result of a number of advantageous features. Without being bound by theory, it is believed in some embodiments, an AAV particle described herein can be used to administer and / or deliver a gene encoding GBA1 (e.g., a human GBA1 protein) preferentially to the CNS (e.g., the brain or the spinal cord).

[0086] Provided herein are compositions and methods which may provide for improved features compared to prior AAV-mediated enzyme replacement approaches, including (i) increasedbiodistribution throughout the CNS (e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region), (ii) elevated GBA1 expression in multiple brain regions (e.g., cortex, thalamus, and brain stem); and (iii) preferential biodistribution in the CNS over the liver and / or DRG. of the subject. In some embodiments, an AAV viral genome comprising a codon-optimized, CpG- reduced (e.g., CpG-depleted) nucleotide sequence encoding a GBA1 protein (e.g.. SEQ ID NO: 2001 or SEQ ID NO: 2002) results in high biodistribution in the CNS; increased GCase activity in the CNS, peripheral tissues, and / or fluid; and successful transgene transcription and expression.

[0087] Also provided herein are AAV capsid variants (e.g., AAV9 capsid variants) with improved properties compared to wildtype AAV9, such as (i) increased penetrance through the blood brain barrier following intravenous administration, (ii) wider distribution throughout the multiple brain regions, e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary' complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region, (iii) elevated GBA1 expression in multiple brain regions, (iv) wider distribution in one or more peripheral tissues, e.g., die heart, kidney, muscle (e.g., quadriceps muscle), pancreas, and / or liver, and / or (v) elevated GBA1 expression in one or more peripheral tissues. In some embodiments, the AAV capsids described herein enhance the delivery of GBA1 to multiple regions of the brain including, for example, die amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g.. cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region). In some embodiments, the AAV capsids described herein enhance the deliver}' of GBA1 to the spinal cord. In some embodiments, the AAV capsid variants with enhanced brain tropism described herein are capable of significantly increasing GBA1 mRNA expression in the brain. In some embodiments, the AAV capsid variants described herein enhance the delivery of GBA1 to the dentate nucleus. In some embodiments, the AAV capsid variants described herein enhance the delivery of GBA1 to the somatosensory cortex.

[0088] Thus, the compositions and methods described herein can be used in the treatment of GBA1- related disorders. In some embodiments, GBA1 -related disorders are disorders associated with a lack of GBA1 protein expression and / or GCase activity and include neuronopathic (affects the CNS) and non- neuronopathic (affects non-CNS) Gaucher disease (e.g., Type 1 GD (GDI), Type 2 GD (GD2), or Type 3 GD (GD3)), PD associated with at least one mutation in a GBA gene. Parkinson’s disease dementia (PDD), Dementia with Lewy Bodies (DLB), and Lew Body Dementia (LBD). In some embodiments, the disclosure provides an AAV particle comprising one of the AAV capsid variants disclosed herein and an AAV viral genome comprising a codon-optimized, CpG-reduced (e.g.. CpG-depleted) nucleotide sequence encoding a GBA1 protein (e.g., comprising the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical to SEQ ID NO: 2001 or SEQ ID NO: 2002). In some embodiments, the codon-optimized. CpG-reduced e.g.. CpG-depleted) nucleotide sequence has reduced immunogenicity compared to a codon-optimized sequence comprising one or more or all CpG motifs. In some embodiments, the codon-optimized, CpG-reduced e.g., CpG-depleted) nucleotide sequence is suitable for use in treating disorders associated with a lack of a GBA1 protein expression and / or GCase activity, such as neuronopathic (affects the CNS) and non -neuronopathic (affects non-CNS). Gaucher disease (e.g., GDI, GD2, or GD3). PD associated with at least one mutation in a GBA gene, Parkinson’s disease dementia (PDD), Dementia with Lewy Bodies (DLB), and I.ewy Body Dementia (LBD).I. CompositionsAdeno-associated viral (AA V) Particles

[0089] AAVs have a genome of about 5,000 nucleotides in length and contains two open reading frames encoding the proteins responsible for replication (Rep) and the structural protein of the capsid (Cap). The open reading frames are flanked by two Inverted Terminal Repeat (ITR) sequences, which serve as the origin of replication of the viral genome. The wild-type AAV viral genome comprises nucleotide sequences for two open reading frames, one for the four non-structural Rep proteins (Rep78, Rep68, Rep52, Rep40. encoded by Rep genes) and one for the three capsid, or structural, proteins (VP1, VP2, VP3, encoded by capsid genes or Cap genes). The Rep proteins are important for replication and packaging, while the capsid proteins are assembled to create the protein shell of the AAV. or AAV capsid. Alternative splicing and alternate initiation codons and promoters result in the generation of four different Rep proteins from a single open reading frame and the generation of three capsid proteins from a single open reading frame. Though it varies by AAV seroty pe, as a non-limiting example, for AAV9 / hu. l 4 (SEQ ID NO: 123 of US 7,906,11 1, the contents of which are herein incorporated by reference in their entirety), VP1 refers to amino acids 1-736, VP2 refers to amino acids 138-736, and VP3 refers to amino acids 203-736. In some embodiments, with reference to the amino acid sequence of SEQ ID NO: 981, VP1 comprises amino acids 1-736, VP2 comprises amino acids 138-736, and VP3 comprises amino acids 203-736. In other words, VP1 is the full-length capsid protein sequence, while VP2 and VP3 are shorter components of the whole. As a result, changes in the sequence in the VP3region are also changes to VP1 and VP2, however, the percent difference as compared to the parent sequence will be greatest for VP3 since it is die shortest sequence of the three. Though described here in relation to the amino acid sequence, the nucleic acid sequence encoding these proteins can be similarly described. Together, the three capsid proteins assemble to create the AAV capsid. Without being bound by theory, the AAV capsid typically comprises a molar ratio of 1: 1: 10 of VP1:VP2:VP3.

[0090] The AAV particle typically requires a co-helper (e.g.. adenovirus) to undergo productive infection in cells. In the absence of such helper functions, the AAV virions essentially enter host cells but do not integrate into the cells’ genome.

[0091] AAV particles have been investigated for delivery of gene therapeutics because of several unique features. Non-limiting examples of the features include (i) the ability to infect both dividing and non-dividing cells: (ii) a broad host range for infectivity, including human cells: (iii) wild-type AAV has not been associated with any disease and has not been shown to replicate in infected cells; (iv) the lack of cell-mediated immune response against the particle, and (v) the non-integrative nature in a host chromosome thereby reducing potential for long-term genetic alterations. Moreover, infection with AAV particles has minimal influence on changing the pattern of cellular gene expression (Stilwell and Samulski el al., Biotechniques, 2003, 34, 148, the contents of which are herein incorporated by reference in their entirety).

[0092] Typically, AAV particles for GBA1 delivery may be recombinant viral particles which are replication defective as they lack sequences encoding functional Rep and Cap proteins within the viral genome. In some cases, the replication defective AAV particles may lack most or all coding sequences and essentially only contain one or two AAV ITR sequences and a nucleic acid sequence encoding a GBA1 protein (e.g., human GBA1 protein).

[0093] In some embodiments, the AAV particles of the present disclosure may be introduced into mammalian cells.

[0094] AAV particles may be modified to enhance the efficiency of delivery. Such modified AAV particles of the present disclosure can be packaged efficiently and can be used to successfully infect the target cells at high frequency and with minimal toxicity'.

[0095] In other embodiments, AAV particles of the present disclosure may be used to deliver GBA1 to the central nervous system (see, e.g., U.S. Pat. No. 6,180,613; the contents of which are herein incorporated by reference in their entirety) or to specific tissues of the CNS.

[0096] It is understood that the compositions described herein may have additional conservative or non-essential amino acid substitutions, which do not have a substantial effect on their functions.

[0097] In some embodiments, an AAV capsid variant comprises one or more modifications in loop VIII of AAV9, e.g., at one or more amino acids corresponding to positions 580-599 of VP1 of AAV9. In some embodiments, an AAV capsid variant comprises a substitution at one. two, three, four, five, or all of amino acids corresponding to position(s) 584, 586, 587. 588, 589, and / or 590 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, the term loop (e.g., loop VIII) is usedinterchangeably herein with the term variable region (e.g., variable region VIII), or VR (e.g., VR-VIII). In some embodiments, loop VIII comprises amino acids corresponding to positions 580-599 (e.g., amino acids VATNHQSAQAQAQTGWVQNQ (SEQ ID NO: 5122)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII is present (is located) at amino acids comprising those corresponding to positions 580-599 (e.g., the positions of amino acids VATNHQSAQAQAQTGWVQNQ (SEQ ID NO: 5122)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII comprises amino acids corresponding to positions 582-593 (e.g., the positions of amino acids TNHQSAQAQAQT (SEQ ID NO: 5123)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII comprises amino acids corresponding to positions 587-593 (e.g.. the positions of amino acids AQAQAQT (SEQ ID NO: 4687)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII comprises amino acids corresponding to positions 587-590 (e.g.. the positions of amino acids AQAQ (SEQ ID NO: 5099)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII is present at amino acids comprising those corresponding to positions 587-590 (e.g., the positions of amino acids AQAQ (SEQ ID NO: 5099)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII or variable region VIII (VR- VIII) is as described in DiMattia et al. “Structural Insights into the Unique Properties of the Adeno- Associated Virus Serotype 9,” Journal of Virology’, 12(86):6947-6958 (the contents of which are hereby incorporated by reference in their entirety), e.g., comprising amino acids corresponding to positions 581 - 593 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII is present at amino acids comprising those corresponding to positions 581-593 of the amino acid sequence of SEQ ID NO: 138.

[0098] The AAV particles and payloads of the disclosure may be delivered to one or more target cells, tissues, organs, or organisms. In some embodiments, the AAV particles demonstrate enhanced tropism for a target cell type, tissue, or organ. As a non-limiting example, the AAV particle may have enhanced tropism for cells and tissues of the central or peripheral nervous systems (CNS and PNS, respectively). In some embodiments, an AAV particle may, in addition, or alternatively, have decreased tropism for a cell-type, tissue or organ.

[0099] In some embodiments, AAV particles are used as a biological tool due to a relatively simple structure, their ability to infect a wide range of cells (including quiescent and dividing cells) without integration into the host genome and without replicating, and their relatively benign immunogenic profile. The genome of the virus may be manipulated to contain a minimum of components for the assembly of a functional recombinant virus, or viral particle, which is loaded with or engineered to target a particular tissue and express or deliver a desired pay load.

[0100] In some embodiments, the AAV particle is a recombinant AAV particle. In some embodiments, the wild-type AAV viral genome is a linear, single-stranded DNA (ssDNA) molecule approximately 5,000 nucleotides (nt) in length. In some embodiments, inverted terminal repeats (ITRs) cap the viral genome at both the 5 ’ and the 3 ’ end. providing origins of replication for the viral genome.In some embodiments, an AAV viral genome comprises tw o ITR sequences. In some embodiments, the ITRs have a characteristic T-shaped hairpin structure defined by a sclf-complcnicntary region (145nt in wild-type AAV) at the 5’ and 3’ ends of the ssDNA which form an energetically stable double stranded region. In some embodiments, the double stranded hairpin structures comprise multiple functions including, but not limited to, acting as an origin for DNA replication by functioning as primers for the endogenous DNA polymerase complex of the host viral replication cell.

[0101] AAV particles of the present disclosure may be produced recombinantly and may be based on AAV reference sequences. In addition to single-stranded AAV viral genomes (e.g., ssAAVs), the present disclosure also provides for self-complementary AAV (scAAVs) viral genomes. scAAV viral genomes contain DNA strands that anneal together to form double-stranded DNA. By skipping second strand synthesis. scAAVs allow for rapid expression in the transduced cell. In some embodiments, the AAV particle of the present disclosure is an scAAV. In some embodiments, the AAV particle of the present disclosure is an ssAAV.

[0102] Methods for producing and / or modifying AAV particles are disclosed in the art such as pseudotyped AAV particles (PCT Patent Publication Nos. W0200028004; W0200123001;W02004112727; W02005005610; and W02005072364. the content of each of w hich is incorporated herein by reference in its entirety).

[0103] As described herein, the AAV particles of the disclosure comprising an AAV capsid variant, and a viral genome, have enhanced tropism for a cell-type or a tissue, e.g., a CNS cell-type, region, or tissue.AAV Capsid Variants

[0104] Disclosed herein are AAV particles comprising an AAV capsid variant comprising one or more modifications (e.g., comprising one or more substitutions relative to a wildly pc AAV capsid) for enhanced or improved transduction of a target tissue (e.g., cells, regions, and / or tissues of the CNS and / or PNS). In some embodiments, the peptide (e g., comprising one or more substitutions relative to a wildly pc AAV capsid) is present in VP1, VP2, and / or VP3 proteins of the AAV capsid variant. In some embodiments, the modification (e.g., comprising one or more substitutions relative to a wildtype AAV capsid) is present in VP1, VP2. and VP3 proteins of the AAV capsid variant.

[0105] In some embodiments, the modification (e.g., comprising the one or more substitutions relative to a wildtype AAV capsid) is in loop VIII. In some embodiments, the AAV capsid variant is an AAV9 capsid variant.

[0106] In some embodiments, the one or more substitutions in the AAV capsid may increase distribution of an AAV particle to a cell, region, or tissue of the CNS. The cell of the CNS may be, but is not limited to, neurons (e.g., excitatory, inhibitory, motor, sensory, autonomic, sympathetic, parasympathetic. Purkinje, Betz, etc.), glial cells (e.g., microglia, astrocytes, oligodendrocytes) and / or supporting cells of the brain such as immune cells (e.g., T cells). The tissue of the CNS may be, but is not limited to, amygdala, brainstem, caudate, central grey, cerebellum (e.g.. Purkinje cell layer and deepcerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g.. cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region.

[0107] In some embodiments, the one or more substitutions may increase distribution of an AAV particle to a cell, region, or tissue of the CNS. In some embodiments, the one or more substitutions may decrease distribution of an AAV particle to the DRG.

[0108] In some embodiments, the one or more substitutions may increase distribution of an AAV particle to the CNS (e.g.. the cortex) after intravenous administration. In some embodiments, the one or more substitutions may increase distribution of an AAV particle to the CNS (e.g.. amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g.. frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region) following focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI- guided FUS coupled with intravenous administration.

[0109] In some embodiments, the one or more substitutions may increase distribution of an AAV particle to the PNS (e.g., DRG) after intravenous administration. In some embodiments, the one or more substitutions may increase distribution of an AAV particle to non-DRG cells of the PNS following focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration. In some embodiments, the one or more substitutions may decrease distribution of an AAV particle to the DRG following focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration.

[0110] In some embodiments, tire one or more modifications, e.g., substitutions, may increase distribution of an AAV particle to a cell, region, or tissue of a heart, e.g., a heart atrium or a heart ventricle. In some embodiments, the one or more substitutions may increase distribution of an AAV particle to a heart cell, region, or tissue after intravenous administration.[oni] In some embodiments, the one or more modifications, e.g., substitutions, may increase distribution of an AAV particle to a cell, region, or tissue of a muscle. In some embodiments, the muscle is a heart muscle (e.g., a heart atrium or a heart ventricle) or a quadriceps muscle. In some embodiments, the one or more substitutions may increase distribution of an AAV particle to a muscle cell, region, or tissue after intravenous administration.

[0112] In some embodiments, the one or more modifications, e.g., substitutions, may increase distribution an AAV particle to a cell, region, or tissue of the kidney. In some embodiments, the one or more substitutions may increase distribution an AAV particle to a cell, region, or tissue of the pancreas.

[0113] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) a sequence as set forth in Table 1. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) a sequence as set forth in Table 2A. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) a sequence as set forth in Table 2B (e.g., a sequence of any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238. 255, 262-265, 274, 283, 286. 290, 291, 293. 301, 306, 307, 308, 309, 314. and 336). In some embodiments, the AAV capsid variant comprises (e.g.. in loop VIII) a sequence set forth in Table 14. In some embodiments, the AAV capsid variant comprises (e.g.. in loop VIII) a sequence as set forth in Table 27. In some embodiments, the AAV capsid variant comprises (e.g.. in loop VIII) a sequence as set forth in Table 28. In some embodiments, the AAV capsid variant is an AAV9 capsid variant.Table 1. Exemplary SequencesTable 2A. Exemplary SequencesTable 2B. Exemplary Sequences

[0114] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence having the formula [N1]-[N2]-[N3] (SEQ ID NO: 4681), wherein [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682) and wherein [Nl] comprises amino acids X X2, X3, and X4, wherein X4 is Q, K, E, S, P, R, N, H, or a conservative substitution thereof and / or [N3] comprises X5. X6, and X7, wherein X5is I, V, T. M, S, N, L, F, or a conservative substitution thereof. In some embodiments, [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682), [Nl] comprises Xi, X2, X3, and X4, wherein X4is Q, K, E, S, P, R. N, or H, and / or [N3] comprises X5. X6, and X7, wherein X5 is I, V. T, M. S, N, L. or F. In some embodiments, X4 of [Nl] is Q. In some embodiments, X4 of [Nl] is K. In some embodiments, X5 of [N3] is I. In some embodiments, Xi of [Nl] is T, S, R. A, I, C.N, K, L, or Q. In some embodiments, X2of [Nl] is N, T, G, V, S, Y, K, I, H, D, or F. In some embodiments, X3of [Nl] is T, N, K, D, I, S, P, A, Y, E, V, L, M, R, H, Q, or C. In some embodiments, [Nl] is or comprises TNTQ (SEQ ID NO: 4688). In some embodiments, [Nl] is or comprises TNTK (SEQ ID NO: 4689). In some embodiments, [N1]-[N2] is or comprises TNTQDWHR (SEQ ID NO: 4898). In some embodiments, [N1]-[N2] is or comprises TNTKDWHR (SEQ ID NO: 4899). In some embodiments, X6of [N3] is A, Y, P, N, S, T, G, E, V, W, F, or Q. In some embodiments, X7of [N3] is Q, G, N, K, H, R. E, L, P, or M. In some embodiments, [N3] is or comprises IAQ. In some embodiments, [N2]-[N3] is or comprises DWHRIAQ (SEQ ID NO: 5027). In some embodiments, [N1]-[N2]-[N3] is or comprises TNTQDWHRIAQ (SEQ ID NO: 343). In some embodiments, [N1]-[N2]-[N3] is or comprises TNTKDWHRIAQ (SEQ ID NO: 344).

[0115] In some embodiments, the amino acid sequence having the formula [N1]-[N2]-[N3] further comprises [N4] . wherein [N4] comprises amino acids X8, X9, X10, and Xn, wherein X8is T, S. N, P, A. or I; X9is G. N, D. R, V. A, S, or Q; X10 is W. S. C, R. L. or G; and / or Xu is V, A. S, I, C, G, D, F. L, or T. In some embodiments, [N4] is or comprises TGWV (SEQ ID NO: 5066). In some embodiments, [Nl ]- [N2]-[N3]-[N4] is or comprises any one of SEQ ID NOs: 201-245, 247-250. 253-255, 257-265. 268-274, 276-286. 288, 290-297. 299-303, 305-309. 311, 313-319. 323-328, 330-337. 339-342, 539-542. 544, 546. 547. 549-557, 559-589. 592, 593, 595, 596, 598. 599, 601-608, 610-614, 616-622. 625, 628, 630. 631, 633. 636, 638, 639-646, 649, 651-657, 667, 669. 670, 672, 673. 679-683, 685-690. 692, 693, 695, 697, 699-701, 703-705, 708-710, 712-717, 719-723, 728-731, 733-738, 740, and 742. In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises TNTQDWHRIAQTGWV (SEQ ID NO: 201). In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises TNTKDWHRIAQTGWV (SEQ ID NO: 202).

[0116] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence having the formula [N1]-[N2]-[N3] (SEQ ID NO: 4683), wherein [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682) and wherein [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4is Q, P, or a conservative substitution thereof; and / or [N3] comprises amino acids X5, X6, and X?, wherein X3is I, V, or a conservative substitution thereof. In some embodiments, [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682); [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4is Q or P; and / or [N3] comprises amino acids X5. Xe, and X7, wherein X, is I or V. In some embodiments, X4of [Nl] is Q. In some embodiments, X, of [N3] is I. In some embodiments, X5 of [N3] is V. In some embodiments, Xi of [Nl] is T or S. In some embodiments, X2of [Nl] is N, T, G, S, I, or V. In some embodiments, X3of [Nl] is T, N, I, S, A, V, or L. In some embodiments, [Nl] is or comprises TNTQ (SEQ ID NO: 4688). In some embodiments, [N1]-[N2] is or comprises TNTQDWHR (SEQ ID NO: 4898). In some embodiments, Xe of [N3] is A, P, S. Y, or N. In some embodiments, X7 of [N3] is Q. G, or N. In some embodiments. [N3] is or comprises IAQ. In some embodiments. [N2]-[N3] is or comprises DWHRIAQ (SEQ ID NO: 5027). In some embodiments, [N 1 ]-[N2]-[N3] is or comprises TNTQDWHRIAQ (SEQ ID NO: 343). In some embodiments, the amino acid sequence having the formula [N1]-[N2]-[N3] further comprises [N4], wherein [N4] comprises amino acids X8. X9, X10. andXii, wherein Xio is W. In some embodiments, Xxof [N4] is T, S, or N. In some embodiments, X9of [N4] is G or N. In some embodiments, Xu of [N4] is V, A, I, or S. In some embodiments, [N4] is or comprises TGWV (SEQ ID NO: 5066). In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291, 293, 301, 306, 307, 308, 309, 314, and 336. In some embodiments, [N1]-[N2]-[N3]-[N4] is or comprises TNTQDWHRIAQTGWV (SEQ ID NO: 201).

[0117] In some embodiments, [N1]-[N2]-[N3] is present in loop VIII of the AAV capsid variant. In some embodiments, [N4] is present in loop VIII of the AAV capsid variant. In some embodiments. [Nl]- [N2]-[N3]-[N4] is present in loop VIII of the AAV capsid variant. In some embodiments, loop VIII is present at amino acids comprising those corresponding to positions 581-593 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, loop VIII is present at amino acids comprising those corresponding to positions 580-599 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981.

[0118] In some embodiments, [Nl] is present at amino acids corresponding to positions 582-585 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, Xi of [Nl] is present at an amino acid corresponding to position 582 of the amino acid sequence of SEQ ID NO: 138 or 981, X2of [Nl] is present at an amino acid corresponding to position 583 of the amino acid sequence of SEQ ID NO: 138 or 981. X3of [Nl] is present at an amino acid corresponding to position 584 of the amino acid sequence of SEQ ID NO: 138 or 981, and X, of [Nl] is present at an amino acid corresponding to position 585 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, [Nl] replaces amino acids at positions corresponding to positions 582-585 (e.g., T582. N583. H584, and Q585) of the ammo acid sequence of SEQ ID NO: 138.

[0119] In some embodiments, [N2] is present at amino acids corresponding to positions 586-589 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, [N2] replaces amino acids at positions corresponding to positions 586-589 of the amino acid sequence of SEQ ID NO: 138 (e.g., S586, A587, Q588, and A589 may be replaced with D586, W587, H588, and R589).

[0120] In some embodiments, [N1]-[N2] is present at ammo acids corresponding to positions 582- 589 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, [Nl]- [N2] replaces amino acids at positions corresponding to positions 582-589 of the amino acid sequence of SEQ ID NO: 138 (e.g.. T582, N583, H584, Q585, S586, A587. Q588, and A589 may be replaced with T582. N583, T584, Q585, D586, W587, H588, and R589).

[0121] In some embodiments, [N3] is present at amino acids corresponding to positions 590-592 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, X5of [N3] is present at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138 or 981, X6of [N3] is present at an amino acid corresponding to position 591 of the amino acid sequence of SEQ ID NO: 138 or 981, and X7of [N3] is present at an amino acid corresponding to position 592 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, [N3] replaces amino acidsat positions corresponding to positions 590-592 (e.g., 1590, A591, and Q592) of die amino acid sequence of SEQ ID NO: 138.

[0122] In some embodiments, [N2]-[N3] is present at amino acids corresponding to positions 586- 592 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, [N2]- [N3] replaces amino acids at positions corresponding to positions 586-592 of the amino acid sequence of SEQ ID NO: 138 (e.g.. S586, A587, Q588, A589, Q590, A591, and Q592 may be replaced with D586, W587, H588, R589, 1590, A591, and Q592).

[0123] In some embodiments, [N1]-[N2]-[N3] is present at amino acids corresponding to positions 582-592 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, [N1]-[N2]-[N3] replaces amino acids at positions corresponding to positions 582-592 of the amino acid sequence of SEQ ID NO: 138 (e.g.. T582. N583, H584, Q585, S586, A587. Q588, A589, Q590, A591. and Q592 may be replaced with T582, N583, T584, Q585. D586, W587, H588. R589. 1590, A591. and Q592).

[0124] In some embodiments, [N4] is present at amino acids corresponding to positions 593-596 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, X§ of [N4] is present at an amino acid corresponding to position 593 of the amino acid sequence of SEQ ID NO: 138 or 981, X9of [N4] is present at an amino acid corresponding to position 594 of the amino acid sequence of SEQ ID NO: 138 or 981. Xin of [N4] is present at an amino acid corresponding to position 595 of the amino acid sequence of SEQ ID NO: 138 or 981, and Xu of [N4] is present at an amino acid corresponding to position 596 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, [N4] replaces amino acids at positions corresponding to positions 593-596 (e.g., T593. G594. W595, and V596) of the amino acid sequence of SEQ ID NO: 138.

[0125] In some embodiments, [N2]-[N3]-[N4] is present at amino acids corresponding to positions 586-596 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, [N2]-[N3]-[N4] replaces amino acids at positions corresponding to positions 586-596 of the amino acid sequence of SEQ ID NO: 138 (e.g., S586, A587, Q588, A589, Q590, A591, Q592, T593, G594, W595, and V596 may be replaced with D586, W587, H588, R589, 1590, A591, Q592, T593, G594, W595, and V596).

[0126] hi some embodiments, [N1]-[N2]-[N3]-[N4] is present at amino acids corresponding to positions 582-596 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, [N1]-[N2]-[N3]-[N4] replaces amino acids at positions corresponding to positions 582-596 of the amino acid sequence of SEQ ID NO: 138 (e.g., T582, N583, H584. Q585, S586, A587, Q588, A589, Q590, A591, Q592, T593, G594. W595, and V596 may be replaced with T582. N583, T584, Q585, D586, W587, H588. R589, 1590. A591, Q592, T593, G594, W595, and V596).

[0127] In some embodiments, [N2] is present immediately subsequent to [Nl], In some embodiments. [N3] is present immediately subsequent to [N2], In some embodiments. [N4] is present immediately subsequent to [N3] . In some embodiments, loop VIII of the AAV capsid variant comprisesan amino acid sequence comprising, from N-terminus to C-tenninus, [N1]-[N2]-[N3], In some embodiments, loop VIII of the AAV capsid variant comprises an amino acid sequence comprising, from N-terminus to C-terminus, [N1]-[N2]-[N3]-[N4],

[0128] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 consecutive amino acids from any one of the sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least 3. at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13. or at least 14 consecutive amino acids from any one of SEQ ID NOs: 201. 205-209, 211-214. 216, 219, 220, 230, 232. 237, 238, 255. 262-265, 274, 283, 286. 290, 291, 293. 301, 306. 307, 308, 309. 314, and 336.

[0129] In some embodiments, the at least 3 consecutive amino acids comprise TQD. In some embodiments, the at least 4 consecutive amino acids comprise TQDW (SEQ ID NO: 4684). In some embodiments, the at least 5 consecutive amino acids comprise TQDWH (SEQ ID NO: 4685). In some embodiments, the at least 6 consecutive amino acids comprise TQDWHR (SEQ ID NO: 4686). In some embodiments, the at least 7 consecutive amino acids comprise TQDWHRI (SEQ ID NO: 941).

[0130] In some embodiments, the at least 3 consecutive amino acids comprise TNT. In some embodiments, the at least 4 consecutive amino acids comprise TNTQ (SEQ ID NO: 4688). In some embodiments, the at least 5 consecutive amino acids comprise TNTQD (SEQ ID NO: 5119). In some embodiments, the at least 6 consecutive amino acids comprise TNTQDW (SEQ ID NO: 5120). In some embodiments, the at least 7 consecutive amino acids comprise TNTQDWH (SEQ ID NO: 5121). In some embodiments, the at least 8 consecutive amino acids comprise TNTQDWHR (SEQ ID NO: 4898). In some embodiments, the at least 9 consecutive amino acids comprise TNTQDWHRI (SEQ ID NO: 746).

[0131] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least one, at least two, or at least three modifications, but no more than four modifications, relative to the amino acid sequence of any one of the sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least one, at least two, or at least three substitutions, but no more than four substitutions, relative to the amino acid sequence of any one of the sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least one. at least two. or at least three modifications, but no more than four modifications, relative to the amino acid sequence of any one of SEQ ID NOs: 201, 205-209, 211-214, 216, 219, 220, 230, 232, 237, 238. 255, 262-265. 274, 283, 286. 290, 291, 293. 301, 306, 307, 308, 309, 314. and 336. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least one, at least two, or at least three substitutions, but no more than four substitutions, relative to the amino acid sequence of any one of SEQ ID NOs: 201, 205-209. 211-214, 216, 219, 220, 230, 232, 237, 238, 255, 262-265, 274, 283, 286, 290, 291, 293, 301, 306, 307, 308, 309, 314, and 336.

[0132] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least one. at least two. or at least three modifications, but no more than four modifications, relative to the amino acid sequence of TQDWHRI (SEQ ID NO: 941). In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence comprising at least one, at least two, or at least three substitutions, but no more than four substitutions, relative to the amino acid sequence of TQDWHRI (SEQ ID NO: 941).

[0133] In some embodiments, the AAV capsid variant comprises the amino acid sequence of any one of the sequences provided in Table 1. 2A. 2B. 14, 27, 28, or 29. In some embodiments, the AAV capsid variant comprises the amino acid sequence of any one of SEQ ID NOs: 201, 205-209. 211-214, 216, 219. 220, 230, 232. 237, 238, 255. 262-265, 274. 283, 286, 290. 291, 293. 301, 306, 307. 308, 309, 314. and 336.

[0134] In some embodiments, the amino acid sequence is present in loop VIII. In some embodiments, loop VIII comprises amino acids corresponding to positions 581-593 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII comprises amino acids corresponding to positions 580-599 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, loop VIII is present (is located) at amino acids comprising those corresponding to positions 580-599 (e.g., amino acids VATNHQSAQAQAQTGWVQNQ (SEQ ID NO: 5122)) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence replaces 1, 2, 3. 4, 5, 6, 7, 8. 9, 10, 11, 12, 13, 14, or all of amino acids corresponding to position(s) 582 (e.g., T582), 583 (e.g., N583), 584 (e.g., H584), 585 (e.g., Q585), 586 (e.g., S586), 587 (e.g., A587), 588 (e.g.. Q588), 589 (e.g., A589). 590 (e.g., Q590), 591 (e.g., A591), 592 (e.g., Q592), 593 (e.g., T593), 594 (e.g., G594), 595 (e.g., W595), and / or 596 (e.g., V596) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence is present at 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or all of amino acids corresponding to position(s) 582, 583, 584, 585, 586, 587, 588, 589, 590, 591, 592, 593, 594, 595, and / or 596 of tire amino acid sequence of SEQ ID NO: 981. In some embodiments, the AAV capsid variant comprises one or more substitutions at amino acids corresponding to position(s) 582 (e.g., T582), 583 (e.g., N583), 584 (e.g., H584), 585 (e.g., Q585), 586 (e.g., S586), 587 (e.g., A587), 588 (e.g., Q588), 589 (e.g., A589), 590 (e.g., Q590), 591 (e.g., A591), 592 (e.g., Q592), 593 (e.g., T593), 594 (e.g., G594), 595 (e.g., W595), and / or 596 (e.g.. V596) of the amino acid sequence of SEQ ID NO: 138.

[0135] In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence encoded by a nucleotide sequence described herein, e.g., a nucleotide sequence of Table 2A. In some embodiments, the AAV capsid variant comprises (e.g.. in loop VIII) an amino acid sequence encoded by a nucleotide sequence comprising at least one. at least two, at least three, at least four, at least five, at least six. or at least seven modifications, but no more than ten modifications, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the AAV capsid variant comprises (e.g.,in loop VIII) an amino acid sequence encoded by a nucleotide sequence comprising at least one, at least tw o. at least three, at least four, at least five, at least six, or at least seven different substitutions, but not more than ten substitutions, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the AAV capsid variant comprises (e.g., in loop VIII) an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 92%, at least 95%. at least 97%, at least 98%, or at least 99% sequence identity) thereto.

[0136] In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, or a portion thereof that is modified relative to a wildtype AAV capsid (e.g., comprising one or more substitutions relative to a wildtype AAV capsid), comprises a nucleotide sequence described herein, e.g., as described in Table 2A. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, or a portion thereof that is modified relative to a wildtype AAV capsid (e.g., comprising one or more substitutions relative to a wildtype AAV capsid), is codon optimized. In some embodiments, the nucleotide sequence is an isolated nucleotide sequence. In some embodiments, the nucleotide sequence is a recombinant nucleotide sequence.

[0137] In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, or a portion thereof that is modified relative to a wildtype AAV capsid (e.g., comprising one or more substitutions relative to a wildtype AAV capsid), comprises the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence comprising at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, but no more than ten modifications, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the nucleotide sequence comprises a nucleotide sequence comprising at least one, at least two, at least three, at least four, at least five, at least six, or at least seven substitutions, but no more than ten substitutions, relative to the nucleotide sequence of SEQ ID NO: 942. In some embodiments, the nucleotide sequence comprises the nucleotide sequence of SEQ ID NO: 942, or a nucleotide sequence substantially identical (e.g., having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity) thereto.

[0138] In some embodiments, the AAV capsid variant further comprises an amino acid other than A at a position corresponding to position 581 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant further comprises the amino acid T at a position corresponding to position 581 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant further comprises the amino acid V at a position corresponding to position 581 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant comprises the substitution A581T or A581V, wherein the substitution is at a position that corresponds to position 581 of the amino acid sequence of SEQ ID NO: 138 or 981.

[0139] In some embodiments, an AAV capsid variant described herein comprises one. two, three, four, five, or all of an amino acid other than H at a position corresponding to position 584 (e.g.,comprises T at an amino acid corresponding to position 584) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than S at a position corresponding to position 586 (e.g., comprises D at an amino acid corresponding to position 586) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than A at a position corresponding to position 587 (e.g., comprises W at an amino acid corresponding to position 587) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than Q at a position corresponding to position 588 (e.g., comprises H at an amino acid corresponding to position 588) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than A at a position corresponding to position 589 (e.g.. comprises R at an amino acid corresponding to position 589) of the amino acid sequence of SEQ ID NO: 138; and / or an amino acid other than Q at a position corresponding to position 590 (e.g.. comprises I at an amino acid corresponding to position 590) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises an amino acid other than H at a position corresponding to position 584 (e.g., comprises T at an amino acid corresponding to position 584) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than S at a position corresponding to position 586 (e.g., comprises D at an amino acid corresponding to position 586) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than A at a position corresponding to position 587 (e.g., comprises W at an amino acid corresponding to position 587) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than Q at a position corresponding to position 588 (e.g., comprises H at an amino acid corresponding to position 588) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than A at a position corresponding to position 589 (e.g., comprises R at an amino acid corresponding to position 589) of the amino acid sequence of SEQ ID NO: 138; and an amino acid other than Q at a position corresponding to position 590 (e.g., comprises I at an amino acid corresponding to position 590) of die amino acid sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein comprises the amino acid T at a position corresponding to position 584 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid D at a position corresponding to position 586 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid W at a position corresponding to position 587 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid H at a position corresponding to position 588 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid R at a position corresponding to position 589 of the amino acid sequence of SEQ ID NO: 138 or 981, and amino acid I at a position corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138 or 981.

[0140] In some embodiments, an AAV capsid variant described herein comprises one. two, three, four, five, or all of the amino acid T at a position corresponding to position 584 of the amino acid sequence of SEQ ID NO: 138 or 981. amino acid D at a position corresponding to position 586 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid W at a position corresponding to position 587 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid H at a position corresponding to position 588 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid R at a position corresponding to position 589 of the amino acid sequence of SEQ ID NO: 138 or 981. and / or amino acidI at a position corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138 or 981. In some embodiments, the AAV capsid variant comprises the amino acid T at a position corresponding to position 584 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid D at a position corresponding to position 586 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid W at a position corresponding to position 587 of the amino acid sequence of SEQ ID NO: 138 or 981. amino acid H at a position corresponding to position 588 of the amino acid sequence of SEQ ID NO: 138 or 981, amino acid R at a position corresponding to position 589 of the amino acid sequence of SEQ ID NO: 138 or 981, and amino acid I at a position corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138 or 981.

[0141] In some embodiments, an AAV capsid variant described herein comprises one. two, three, four, five, or all of the substitutions H584T. S586D, A587W, Q588H, A589R, and / or Q590I, wherein the substitution(s) is / are at position(s) that correspond to position / s) 584. 586, 587, 588. 589, and / or 590 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises the substitutions H584T, S586D, A587W, Q588H, A589R, and Q590I, wherein the substitution(s) is / are at position(s) that correspond to position(s) 584, 586. 587, 588, 589. and / or 590 of the amino acid sequence of SEQ ID NO: 138 or 981.

[0142] In some embodiments, an AAV capsid variant described herein comprises the amino acid Q at an amino acid corresponding to position 585 of the amino acid sequence of SEQ ID NO: 138 or 981.

[0143] In some embodiments, an AAV capsid variant described herein comprises an amino acid other than Q at an amino acid corresponding to position 585 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein comprises the amino acid K at an amino acid corresponding to position 585 of the amino acid sequence of SEQ ID NO: 138.

[0144] In some embodiments, an AAV capsid variant described herein comprises an amino acid other than Q at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises the amino acid I at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, die AAV capsid variant comprises the amino acid V at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 138.

[0145] In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941), wherein TQDWHRI (SEQ ID NO: 941) is present in the AAV capsid variant at amino acids corresponding to positions 584-590 of the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. In some embodiments, TQDWHRI (SEQ ID NO: 941) is present at amino acids corresponding to positions 584-590 of the amino acid sequence of SEQ ID NO: 138 (e.g., H584, S586, A587, Q588, A589, and Q590 may be replaced with T584, D586, W587. H588, R589, and 1590).

[0146] In some embodiments, an AAV capsid variant described herein comprises the amino acid W at an amino acid corresponding to position 595 of the amino acid sequence of SEQ ID NO: 138 or 981.

[0147] In some embodiments, the AAV capsid variant further comprises a substitution corresponding to amino acid K449 (e.g., K449R) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises an amino acid other than K at a position corresponding to position 449 (e.g., comprises an R corresponding to position 449) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises an R at amino acid corresponding to position 449 of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises a modification in loop I, II, IV. and / or VI.

[0148] In some embodiments, the AAV capsid variant further comprises an amino acid sequence comprising at least one, at least two, or at least three modifications, but not more than 30, not more than 20, or not more than 10 modifications, relative to the amino acid sequence of SEQ ID NO: 138. In some embodiments, the amino acid sequence does not comprise more than 10 modifications. In some embodiments, the AAV capsid variant further comprises an amino acid sequence comprising at least one. at least two. or at least three, but not more than 30, not more than 20. or not more than 10 amino acids that differ from the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant further comprises the amino acid sequence of SEQ ID NO: 138, or an amino acid sequence with at least 70% (e.g., at least 70%, at least 75%. at least 80%, at least 85%. at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto.

[0149] In some embodiments, the AAV capsid variant comprises (a) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 981 (i.e.. comprising amino acids 1-736 of SEQ ID NO: 981); (b) a VP2 protein comprising amino acids 138-736 of SEQ ID NO: 981; (c) a VP3 protein comprising amino acids 203-736 of SEQ ID NO: 981; or (d) an amino acid sequence with at least 70% (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity to any one of the amino acid sequences in (a)-(c), an amino acid sequence comprising at least one, at least tw o, or at least three substitutions, but not more than 30, not more than 20, or not more than 10 substitutions relative to any one of tire amino acid sequences in (a)-(c), or an amino acid sequence comprising at least one, at least tw o. or at least three modifications, but not more than 30, not more than 20, or not more than 10 such modifications relative to any one of the amino acid sequences in (a)-(c).

[0150] In some embodiments, the AAV capsid variant comprises (a) a VP1 protein comprising an amino acid sequence that is at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 981; (b) a VP2 protein comprising an amino acid sequence that is at least 90% (e.g.. at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to amino acids 138-736 of SEQ ID NO: 981; and / or (c) a VP3 protein comprising an amino acid sequence that is at least 90% (e.g.. at least 90%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99%) identical to amino acids 203-736 of SEQ ID NO: 981, wherein the AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941) or a sequence comprising one. two, or three but not more than three substitutions relative to theamino acid sequence of TQDWHRI (SEQ ID NO: 941). In some embodiments, the AAV capsid variant comprises the amino acid sequence comprising one, tw o, or three but not more than three substitutions relative to the amino acid sequence of TQDWHRI (SEQ ID NO: 941) in loop VIII. In some embodiments, the AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941) in loop VIII.

[0151] In some embodiments, the AAV capsid variant comprises an amino acid sequence as described herein, e.g., an amino acid sequence of TTJ-001. e.g., as described in Tables 3 and 4. In some embodiments, the AAV capsid variant is or comprises TTJ-001. In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 941 in a loop VIII region.

[0152] In some embodiments, the AAV capsid variant comprises a VP1, VP2, and / or VP3 protein comprising an amino acid sequence described herein, e.g., an amino acid sequence of TTJ-001, e.g., as described in Tables 3 and 4.

[0153] In some embodiments, the AAV capsid variant comprises an amino acid sequence encoded by a nucleotide sequence as described herein, e.g., a nucleotide sequence encoding TTJ-001, e.g., as described in Tables 3 and 5.

[0154] In some embodiments, the polynucleotide or nucleic acid encoding the AAV capsid variant comprises a nucleotide sequence described herein, e.g.. a nucleotide sequence encoding TTJ-001. e.g., as described in Tables 3 and 5.Table 3. Exemplary full length capsid sequencesTable 4. Exemplary full length capsid amino acid sequencesTable 5. Exemplary full length capsid nucleic acid sequences

[0155] In some embodiments, the polynucleotide encoding an AAV capsid variant comprises the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence with at least 70% (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto.

[0156] In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, comprises a nucleotide sequence comprising at least one, at least two, or at least three modifications but not more than 30, not more than 20, or not more than 10 modifications, relative to the nucleotide sequence of SEQ ID NO: 983. In some embodiments, the nucleotide sequence encoding an AAV capsid variant described herein, comprises a nucleotide sequence comprising at least one, at least two, or at least three substitutions, but not more than 30. not more than 20, or not more than 10substitutions relative to the amino acid sequence of SEQ ID NO: 983. In some embodiments, the nucleic acid sequence encoding an AAV capsid variant described herein is codon optimized.

[0157] In some embodiments, the AAV capsid variant comprises one or more substitutions in loop VIII and comprises the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence with at least 70% (e.g.. at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one. at least two. or at least three modifications, but not more than 30. not more than 20, or not more than 10 modifications, relative to the amino acid sequence of SEQ ID NO: 981. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence comprising at least one. at least two. or at least three substitutions, but not more than 30, not more than 20, or not more than 10 substitutions, relative to the amino acid sequence of SEQ ID NO: 981.

[0158] In some embodiments, the AAV capsid variant comprises one or more substitutions in loop VIII and comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983, or a nucleotide sequence with at least 70% (e.g., at least 70%, at least 75%. at least 80%, at least 85%. at least 90%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99%) sequence identity thereto. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, at least two, or at least three substitutions, but not more than 30, not more than 20, or not more than 10 substitutions, relative to the amino acid sequence of SEQ ID NO: 983. In some embodiments, an AAV capsid variant described herein comprises an amino acid sequence encoded by a nucleotide sequence comprising at least one, at least two, or at least three modifications, but not more than 30, not more than 20, or not more than 10 modifications, relative to the nucleotide sequence of SEQ ID NO: 983.

[0159] In some embodiments, an AAV capsid variant described herein comprises a VP1, VP2, VP3 protein, wherein at least one of the VP1, VP2, and VP3 comprise one or more substitutions in loop VIII. In some embodiments, the AAV capsid variant comprises amino acids 138-736, e.g., a VP2, of the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence with at least 70% (e.g., at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto. In some embodiments, the AAV capsid protein comprises amino acids 203-736, e.g., a VP3, of the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence with at least 70% (e.g., at least 70%. at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto. In some embodiments, the AAV capsid variant comprises amino acids 1-736, e.g.. a VP1, of the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence with at least 70% (e.g., at least 70%. at least 75%, at least 80%. at least 85%, at least 90%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99%) sequence identity thereto.

[0160] In some embodiments, an AAV capsid variant described herein comprises the amino acid sequence of amino acids 203-736 of SEQ ID NO: 981, or an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto. In some embodiments, the AAV capsid variant comprises the amino acid sequence of amino acids 138-736 of SEQ ID NO: 981, or an amino acid sequence that is at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical thereto. In some embodiments, the AAV capsid variant comprises the amino acid sequence SEQ ID NO: 981, or an amino acid sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical thereto.

[0161] In some embodiments, an AAV capsid variant, described herein has an increased tropism for a CNS cell or tissue, e.g., a brain cell, brain tissue, spinal cord cell, or spinal cord tissue, relative to the tropism of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138.

[0162] In some embodiments, an AAV capsid variant described herein transduces a brain region, e.g.. a midbrain region (e.g., the hippocampus, or thalamus) or the brain stem. In some embodiments, the level of transduction is at least 39. at least 50, at least 100, at least 120, at least 132, at least 146, at least 150. at least 161. at least 174. at least 175. at least 200. at least 225. at least 250. at least 275, at least 283, at least 300, at least 350, at least 400, at least 450, at least 500, at least 525, at least 528, or at least 550- fold greater as compared to an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138.

[0163] In some embodiments, an AAV capsid variant described herein is enriched at least 10, at least 14, at least 20, at least 24, at least 50, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 425, at least 450, or at least 460-fold in the brain compared to enrichment in the brain of an AAV capsid comprising tire amino acid sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein is enriched at least 200, at least 250, at least 300, at least 350, at least 400, at least 425, at least 450, or at least 460-fold in the brain compared to enrichment in the brain of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138.

[0164] In some embodiments, an AAV capsid variant described herein is enriched in the brain of at least two to three species, e.g., a non-human primate and rodent (e.g., mouse) species, compared to enrichment in the brain of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, an AAV capsid variant described herein is enriched at least 2, at least 3, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35. at least 40, at least 45, at least 50, at least 55, at least 60, at least 65. at least 70, at least 75, at least 80. at least 85, at least 90, at least 95, at least 100, at least 105, at least 115, at least 120, at least 125, at least 130, at least 135, at least 140, at least 145, at least 150, at least 155, at least 160, at least 165, at least 170, at least 175, at least 180, at least 190. at least 200. at least 205. or at least 210-fold in the brain of at least two to three species, e.g.. a non- human primate and rodent (e.g., mouse) species, compared to enrichment in the brain of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the at least two to threespecies are Macaca fascicularis, Chlorocebus sabaeus, Callithrix jacchus, and / or mouse (e.g., outbred mice).

[0165] In some embodiments, an AAV capsid variant described herein is enriched at least 2, at least 3. at least 4, at least 5, at least 10. at least 15, at least 17, at least 20, at least 50, at least 75, at least 100, at least 103, at least 107, at least 125, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, or at least 1200-fold in the brain compared to enrichment in the brain of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 981.

[0166] In some embodiments, an AAV capsid variant described herein delivers an increased level of viral genomes to a brain region. In some embodiments, the level of viral genomes is increased by at least 2. at least 5. at least 7, at least 10. at least 15, at least 19. at least 20, at least 22, or at least 25-fold, as compared to the level of viral genomes delivered by an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the brain region comprises a sensory cortex, motor cortex, putamen, thalamus, caudate, hippocampus, and / or cerebellum.

[0167] In some embodiments, an AAV capsid variant delivers an increased level of a payload to a brain region. In some embodiments, the level of the payload is increased by at least 39. at least 50, at least 100, at least 120, at least 132, at least 146, at least 150, at least 161, at least 174, at least 175, at least 200. at least 225. at least 250. at least 275. at least 283. at least 300. at least 350. at least 400, at least 450, at least 500, at least 525, at least 528, or at least 550-fold, as compared to the level of a payload delivered by an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138. In some embodiments, the brain region comprises a sensory cortex, motor cortex, putamen, thalamus, caudate, hippocampus, and / or cerebellum.

[0168] In some embodiments, an AAV capsid variant described herein is enriched at least 5, at least 10, at least 50, at least 100, at least 115, at least 120, at least 150, at least 175, at least 200, at least 207, at least 225, at least 250, or at least 275-fold in the spinal cord compared to enrichment in the spinal cord of an AAV capsid comprising the amino acid sequence of SEQ ID NO: 138.

[0169] In some embodiments, an AAV capsid variant of the present disclosure has decreased tropism for the liver. In some embodiments, an AAV capsid variant comprises a modification that results in reduced tropism (e.g., de-targeting) and / or activity in the liver. In some embodiments, the reduced tropism in the liver is compared to an otherwise similar capsid that does not comprise the modification, e.g., a wild-type capsid polypeptide. In some embodiments, an AAV capsid variant comprises a modification that results in one or more of the following properties: (1) reduced tropism in the liver; (2) de-targeted expression in the liver; (3) reduced activity in the liver; and / or (4) reduced binding to galactose. In some embodiments, the reduction in any one or all of properties (l)-(3) is compared to an otherwise similar AAV capsid variant that does not comprise the modification. Exemplary modifications are provided in WO 2018 / 119330; Pulicherla et al. (2011) Mol. Ther. 19(6): 1070-1078; Adachi et al. (2014) Nature Communications 5(3075), DOI: 10.1038 / ncomms4075; and Bell et al. (2012) J. Virol. 86(13): 7326-33; the contents of which are hereby incorporated by reference in their entirety.

[0170] In some embodiments, the AAV capsid variant comprises a modification at an amino acid corresponding to N470 (e.g., comprises N470A at an amino acid corresponding to N470) of the amino acid sequence of SEQ ID NO: 138; an amino acid corresponding to D271 (e.g., comprises D271 A at an amino acid corresponding to D271) of the amino acid sequence of SEQ ID NO: 138; an amino acid corresponding to N272 (e.g., comprises N272A at an amino acid corresponding to N272) of the amino acid sequence of SEQ ID NO: 138; an amino acid corresponding to Y446 (e.g., comprises Y446A at an amino acid corresponding to Y446) of the amino acid sequence of SEQ ID NO: 138; an amino acid corresponding to N498 (e.g., comprises N498Y or N498I at an amino acid corresponding to N498) of the amino acid sequence of SEQ ID NO: 138; an amino acid corresponding to W503 (e.g., comprises W503R or W503A at an amino acid corresponding to W503) of the amino acid sequence of SEQ ID NO: 138; an amino acid corresponding to L620 (e.g.. comprises L620F at an amino acid corresponding to L620) of the amino acid sequence of SEQ ID NO: 138; or a combination thereof. In some embodiments, the AAV capsid variant comprises one. two, three, four, five, or all of an amino acid other than N at a position corresponding to position 470 (e.g., comprises A at an amino acid corresponding to position 470) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than D at a position corresponding to position 271 (e.g., comprises A at an amino acid corresponding to position 271) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than N at a position corresponding to position 272 (e.g., comprises A at an amino acid corresponding to position 272) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than Y at a position corresponding to position 446 (e.g., comprises A at an amino acid corresponding to position 446) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than N at a position corresponding to position 498 (e.g., comprises Y or I at an amino acid corresponding to position 498) of the amino acid sequence of SEQ ID NO: 138; an amino acid other than W at a position corresponding to position 503 (e.g., comprises R or A at an amino acid corresponding to position 503) of the amino acid sequence of SEQ ID NO: 138; and an amino acid other than L at a position corresponding to position 620 (e.g., comprises F at an amino acid corresponding to position 620) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, die AAV capsid variant comprises a modification at positions corresponding to amino acid N470 (e.g., comprises N470A at an amino acid corresponding to N470) of the amino acid sequence of SEQ ID NO: 138; corresponding to amino acid D271 (e.g., comprises D271 A at an amino acid corresponding to D271) of the amino acid sequence of SEQ ID NO: 138; corresponding to amino acid N272 (e.g., comprises N272A at an amino acid corresponding to N272) of the amino acid sequence of SEQ ID NO: 138; corresponding to amino acid Y446 (e.g., comprises Y446A at an amino acid corresponding to Y446) of the amino acid sequence of SEQ ID NO: 138; and corresponding to amino acid W503 (e.g., comprises W503R or W503A at an amino acid corresponding to W503) of the amino acid sequence of SEQ ID NO: 138. In some embodiments, the AAV capsid variant comprises a modification corresponding to amino acid N498 (e.g.. comprises N498Y at an amino acid corresponding to N498) of the amino acid sequenceof SEQ ID NO: 138 and corresponding to amino acid L620 (e.g., comprises L620F at an amino acid corresponding to L620) of the amino acid sequence of SEQ ID NO: 138.

[0171] In some embodiments, the AAV capsid variant comprises a modification as described in Adachi et al. (2014) Nature Communications 5(3075), DOI: 10.1038 / ncomms4075, the contents of which are hereby incorporated by reference in its entirety . Exemplary modifications that alter or do not alter tissue transduction in at least the brain, liver, heart, lung, and / or kidney can be found in Supplementary Data 2 showing the AAV Barcode-Seq data obtained with AAV9-AA-VBCLib of Adachi et al. (supra), the contents of which are hereby incorporated by reference in their entirety.

[0172] In some embodiments, the AAV capsid variant is an isolated capsid variant. In some embodiments, the AAV capsid variant is a recombinant capsid variant. In some embodiments, a polynucleotide encoding an AAV capsid polypeptide, e.g.. an AAV capsid variant is an isolated and / or a recombinant AAV capsid polypeptide.

[0173] Also provided herein are polynucleotide sequences encoding any of the AAV capsid variants described above and AAV particles, vectors, and cells comprising the same.AAV serotypes and capsids

[0174] In some embodiments, the AAV particle may comprise a capsid protein or variant of any natural or recombinant AAV serotype. AAV serotypes may differ in characteristics such as, but not limited to, packaging, tropism, transduction and immunogenic profdes. Without being bound by theory, it is believed in some embodiments that the AAV capsid protein, e.g.. an AAV capsid variant, can modulate AAV particle tropism in a particular tissue.

[0175] In some embodiments, the AAV capsid variant allow s for blood brain barrier penetration following intravenous administration. In some embodiments, the AAV capsid variant allows for blood brain barrier penetration following intravenous administration, focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration. In some embodiments, the AAV capsid variant allows for increased distribution to a brain region. In some embodiments, the brain region comprises the amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and / or deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, visual cortex, and / or temporal cortex), hilus of the dentate gyrus, external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus. oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, and / or vestibular nucleus. In some embodiments, the AAV capsid variant allows for preferential transduction in a brain region relative to the transduction in the dorsal root ganglia (DRG). In some embodiments, the AAV capsid variant allows for preferential transduction in a brain region relative to the transduction in the liver. In some embodiments, the AAV capsid variant allows for transduction in neuronal cells. In some embodiments, the AAV capsid variant allows for transduction in a non-neuronal cell, e.g.. a glial cell (e.g.. an astrocyte, an oligodendrocyte, or a combination thereof). In some embodiments, the AAV capsid variant allows for transduction in bothneuronal cells and non-neuronal cell, e.g., a glial cell (e.g., an astrocyte, an oligodendrocyte, or a combination thereof).

[0176] In some embodiments, an AAV capsid variant allows for increased distribution to a spinal cord region. In some embodiments, the spinal region comprises a cervical spinal cord region, thoracic spinal cord region, and / or lumbar spinal cord region.

[0177] In some embodiments, the AAV capsid variant allows for increased distribution to a heart region.

[0178] In some embodiments, the AAV capsid variant is suitable for intramuscular administration and / or transduction of muscle fibers. In some embodiments, the AAV capsid variant allows for increased distribution to a muscle region. In some embodiments, the muscle region comprises a heart muscle, quadriceps muscle, a diaphragm muscle region, or a combination thereof. In some embodiments, the muscle region comprises a heart muscle region, e.g.. a heart atrium muscle region or a heart ventricle muscle region.

[0179] In some embodiments, the AAV capsid variant is suitable for increased distribution to a kidney. In some embodiments, the AAV capsid variant is suitable for increased distribution to a pancreas.

[0180] In some embodiments, the initiation codon for translation of the AAV VP1 capsid protein, e.g.. a capsid variant, described herein may be CTG, TTG, or GTG as described in US Patent No. US8163543. the contents of which are herein incorporated by reference in its entirety.

[0181] The present disclosure refers to structural capsid proteins (including VP1, VP2 and VP3) which are encoded by capsid (Cap) genes. These capsid proteins form an outer protein structural shell (e.g., capsid) of a viral vector such as AAV. VP capsid proteins synthesized from Cap polynucleotides generally include a methionine as the first amino acid in the peptide sequence (Metl), which is associated with the start codon (AUG or ATG) in the corresponding Cap nucleotide sequence. However, it is common for a first-methionine (Metl) residue or generally any first amino acid (AA1) to be cleaved off after or during polypeptide synthesis by protein processing enzy mes such as Mct-aminopcptidascs. This “Met / AA-clipping” process often correlates with a corresponding acetylation of the second amino acid in the polypeptide sequence (e.g., alanine, valine, serine, threonine, etc.). Met-clipping commonly occurs with VP1 and VP3 capsid proteins but can also occur with VP2 capsid proteins.

[0182] Where the Met / AA-clipping is incomplete, a mixture of one or more (one, two or three) VP capsid proteins comprising the viral capsid may be produced, some of which may include a Metl / AAl amino acid (Met+ / AA+) and some of which may lack a Metl / AAl amino acid as a result of Met / AA- clipping (Met- / AA-). For further discussion regarding Met / AA-clipping in capsid proteins, see Jin, et al. Direct Liquid Chromatography / Mass Spectrometry Analysis for Complete Characterization of Recombinant Adeno- Associated Virus Capsid Proteins. Hum Gene Ther Methods. 2017 Oct. 28(5):255- 267; Hwang, et al. N-Terminal Acetylation of Cellular Proteins Creates Specific Degradation Signals. Science. 2010 February 19. 327(5968): 973-977; the contents of which are each incorporated herein by reference in its entirety7.

[0183] According to the present disclosure, references to capsid proteins, e.g., AAV capsid variants, is not limited to either clipped (Met- / AA-) or unclipped (Met+ / AA+) and may, in context, refer to independent capsid proteins, viral capsids comprised of a mixture of capsid proteins, and / or polynucleotide sequences (or fragments thereof) which encode, describe, produce or result in capsid proteins of the present disclosure. A direct reference to a capsid protein or capsid polypeptide (such as VP1, VP2 or VP2) may also comprise VP capsid proteins which include a Metl / AAl amino acid (Met+ / AA+) as well as corresponding VP capsid proteins which lack the Metl / AAl amino acid as a result of Met / AA-clipping (Met- / AA-).

[0184] Further according to the present disclosure, a reference to a specific SEQ ID NO (whether a protein or nucleic acid) which comprises or encodes, respectively, one or more capsid proteins which include a Metl / AAl amino acid (Met+ / AA+) should be understood to teach the VP capsid proteins which lack the Metl / AAl amino acid as upon review of the sequence, it is readily apparent any sequence which merely lacks the first listed amino acid (whether or not Metl / AAl).

[0185] As a non-limiting example, reference to a VP 1 polypeptide sequence which is 736 amino acids in length and which includes a “Metl” amino acid (Met+) encoded by the AUG / ATG start codon may also be understood to teach a VP1 polypeptide sequence which is 735 amino acids in length and which does not include the “Metl” amino acid (Met-) of the 736 amino acid Met+ sequence. As a second non-limiting example, reference to a VP 1 polypeptide sequence which is 736 amino acids in length and which includes an “AAl” amino acid (AA1+) encoded by any NNN initiator codon may also be understood to teach a VP1 polypeptide sequence which is 735 amino acids in length and which does not include the “AAl” amino acid (AA1-) of the 736 amino acid AA1+ sequence.

[0186] References to viral capsids formed from VP capsid proteins (such as reference to specific AAV capsid serotypes), can incorporate VP capsid proteins which include a Metl / AAl ammo acid (Met+ / AA1+). corresponding VP capsid proteins which lack the Metl / AAl amino acid as a result of Met / AAl -clipping (Mct- / AA1-), and combinations thereof (Mct+ / AA1+ and Mct- / AA1-).

[0187] As a non-limiting example, an AAV capsid serotype can include VP1 (Met+ / AA1+), VP1 (Met- / AA1-), or a combination of VP1 (Met+ / AA1+) and VP1 (Met- / AA1-). An AAV capsid serotype can also include VP3 (Met+ / AA1+), VP3 (Met- / AA1-), or a combination of VP3 (Met+ / AA1+) and VP3 (Met- / AA1-); and can also include similar optional combinations of VP2 (Met+ / AA1) and VP2 (Met- / AA1-).Additional AAV Capsid Sequences

[0188] In some embodiments, the AAV capsid variant comprises at amino acids corresponding to positions 582, 583, 584, 585, 586, 587, 588, 589, and / or 590 of the amino acid sequence of SEQ ID NO: 138. at least 3, at least 4, at least 5, at least 6. at least 7. at least 8, at least 9, at least 10. at least 11, at least12, at least 13, at least 14, or at least 15 consecutive amino acids of any one of the amino acid sequences provided in Table 1, 2A, 2B, 14, 27, 28, or 29.

[0189] In some embodiments, the AAV capsid variant comprises, immediately subsequent to an amino acids corresponding to position 582, 583, 584, 585, 586. 587, 588, 589, and / or 590 of the amino acid sequence of SEQ ID NO: 138 (e.g., corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrhlO, AAVrh32.33. AAVrh74, PHP.N, PHP.B. or an AAV serotype as provided in Table 6 of WO 2021 / 230987 (the contents of which are hereby incorporated by reference in their entirety))), at least 3, at least 4, at least 5, at least 6, at least 7. at least 8, at least 9, at least 10. at least 11, at least 12, at least 13. at least 14, or at least 15 consecutive amino acids of any one of the amino acid sequences provided in Table 1. 2A, 2B. 14. 27, 28, or 29. In some embodiments, the at least 3. at least 4. at least 5, at least 6, at least 7, at least 8, at least 9, at least 10. at least 11, at least 12, at least 13. at least 14, or at least 15 consecutive amino acids of any one of the amino acid sequences provided in Table 1. 2A. 2B. 14, 27, 28. or 29 replaces at least one, at least two. at least three, at least four, at least five, at least six, at least seven, at least eight, or all of amino acid(s) corresponding to position(s) 582. 583. 584, 585. 586, 587, 588. 589, and / or 590 (e.g., T582, N583. H584, Q585, S586, A587, Q588. A589. and / or Q590) of the amino acid sequence of SEQ ID NO: 138 (e.g., corresponding to equivalent positions in any other AAV serotype (e.g., AAV1. AAV2, AAV3, AAV3b. AAV4, AAV6, AAV7, AAV8. AAV9, AAVrh8, AAVrhlO, AAVrh32.33, AAVrh74, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021 / 230987)). In some embodiments, the AAV capsid variant comprises an amino acid other than the wild-type amino acid, at one, two, three, four, five, six, seven, eight, or all of amino acid(s) corresponding to position(s) 582, 583, 584, 585, 586, 587, 588, 589, and / or 590 of the amino acid sequence of SEQ ID NO: 138 (e.g., corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b, AAV4, AAV6, AAV7, AAV8. AAV9, AAVrh8, AAVrhlO, AAVrh32.33, AAVrh74, PHP.N, PHP.B, or an AAV serotype as provided in Table 6 of WO 2021 / 230987). In some embodiments, the AAV capsid variant comprises a modification replacing one, two. three, four, five, six, seven, eight, or all amino acid(s) corresponding to position(s) 582, 583, 584, 585, 586, 587, 588, 589, and / or 590 (e.g., T582, N583. H584, Q585, S586, A587, Q588, A589, and / or Q590) of the amino acid sequence of SEQ ID NO: 138 (e.g., corresponding to equivalent positions in any other AAV serotype (e.g., AAV1, AAV2, AAV3, AAV3b. AAV4, AAV6, AAV7, AAV8. AAV9, AAVrh8, AAVrhlO. AAVrh32.33, AAVrh74. PHP.N. PHP.B, or an AAV serotype as provided in Table 6 of WO 2021 / 230987).

[0190] In some embodiments, the AAV capsid variant may comprise a variant of a VOY101 capsid polypeptide, an AAVPHP.B (PHP.B) capsid polypeptide, a AAVPHP.N (PHP.N) capsid polypeptide, an AAV1 capsid polypeptide, an AAV2 capsid polypeptide, an AAV5 capsid polypeptide, an AAV9 capsid polypeptide, an AAV9 K449R capsid polypeptide, or an AAVrhlO capsid polypeptide. In some embodiments, the AAV capsid polypeptide, e.g., AAV capsid variant, comprises an amino acid sequenceof any one of the AAV capsid polypeptides in Table 6, or an amino acid sequence substantially identical (e.g., having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity ) thereto. In some embodiments, the nucleotide sequence encoding the AAV capsid polypeptide comprises any one of die nucleotide sequences in Table 6, or a nucleotide sequence substantially identical (e.g., having at least 70%, at least 75%, at least 80%. at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 98%, or at least 99% sequence identity) thereto.Table 6. AAV SequencesAAV Viral Genome

[0191] In some embodiments, the AAV particle of the present disclosure serves as an expression vector comprising a viral genome that encodes a GBA1 protein (e.g., a wildtype GBA1 protein, e.g., a wildtype human GBA1 protein).

[0192] In some embodiments, an AAV particle, e.g., an AAV particle for the vectorized deliver}' of GBA1 described herein, comprises a viral genome, e.g., an AAV viral genome, also referred to as an AAV genome, a vector genome, or an AAV vector genome. In some embodiments, tire viral genome, e.g., the AAV viral genome, further comprises an inverted terminal repeat (ITR) region, an enhancer, a promoter, an intron region, an exon region, a nucleic acid encoding a transgene encoding a GBA1 protein, a nucleotide sequence encoding at least one miR binding site (e.g., at least one miR183 binding site), a polyA region, or a combination thereof.Viral Genome Component: Inverted Terminal Repeats (ITRs)

[0193] In some embodiments, the viral genome may comprise at least one inverted terminal repeat (ITR) region. The AAV particles of the present disclosure comprise a viral genome with at least one ITR region and a region comprising a GBAl-encoding sequence. In some embodiments, the viral genome has two ITRs. These two ITRs flank the region comprising the GBAl-encoding sequence at the 5‘ and 3’ ends. In some embodiments, the ITR functions as an origin of replication comprising a recognition sitefor replication. In some embodiments, the ITR comprises a sequence region which can be complementary and symmetrically arranged. In some embodiments, the ITR incorporated into a viral genome described herein may be comprised of a naturally occurring polynucleotide sequence or a recombinantly derived polynucleotide sequence.

[0194] In some embodiments, the ITR is of the same serotype as the capsid, selected from any one of the serotypes known in the art, or a derivative thereof. In some embodiments, the ITR is of a different serotype than the capsid. In some embodiments, the AAV particle has more than one ITR. In some embodiments, the AAV particle comprises a viral genome comprising two ITRs. In some embodiments, the ITRs are of the same serotype as one another. In some embodiments, the ITRs are of different serotypes. Non-limiting examples include zero, one. or both of the ITRs having the same serotype as the capsid. In some embodiments, both ITRs of the viral genome of the AAV particle are AAV2 ITRs.

[0195] Independently, each ITR may be about 100 to about 150 nucleotides in length. In some embodiments, the ITR is about 120-140 nucleotides in length, e.g.. about 130 nucleotides in length.

[0196] In some embodiments, each ITR is 141 nucleotides in length. In some embodiments, each ITR is 130 nucleotides in length. In some embodiments, the AAV particles comprise two ITRs and one ITR is 141 nucleotides in length and the other ITR is 130 nucleotides in length.

[0197] In some embodiments, the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or 1830, or a nucleotide sequence that is at least 70%. at least 75%, at least 80%, at least 85%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 1829 or 1830. In some embodiments, the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or 1830, or a nucleotide sequence having one, two. three, four, but no more than four modifications, e.g., substitutions, relative to any one of SEQ ID NOs: 1829 or 1830. In some embodiments, the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or 1830. In some embodiments, the viral genome comprises a 5’ ITR comprising the nucleotide sequence of SEQ ID NO: 1829 and a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830.Viral Genome Component: Promoters and Enhancers

[0198] In some embodiments, the viral genome comprises at least one element to enhance the transgene target specificity and expression. See, e.g., Powell et al. Viral Expression Cassette Elements to Enhance Transgene Target Specificity and Expression in Gene Therapy, 2015; tire contents of which are herein incorporated by reference in their entirety. Non-limiting examples of elements to enhance the transgene target specificity and expression include promoters, endogenous miRNAs, post-transcriptional regulatory elements (PREs), polyadenylation (poly A) sequences, upstream enhancers (USEs), CMV enhancers, and introns.

[0199] In some embodiments, expression of the polypeptides in a target cell may be driven by a specific promoter, including but not limited to, a promoter that is species specific, inducible, tissuespecific, or cell cycle-specific (Parr et al.. Nat. Med.3'.1145-9 (1997); the contents of which are herein incorporated by reference in their entirety).

[0200] In some embodiments, the viral genome provides expression of a GBA1 protein encoded by a transgene for expression in a target tissue. In some embodiments, the promoter is deemed to be efficient when it drives expression of the GBA1 encoded in the viral genome of the AAV particle.

[0201] In some embodiments, the promoter is a promoter deemed to be efficient when it drives expression in the cell or tissue being targeted (e.g., the CNS).

[0202] Promoters may be naturally occurring or non-naturally occurring. Non-limiting examples of promoters include viral promoters, plant promoters and mammalian promoters. In some embodiments, the promoters may be human promoters. In some embodiments, the promoter may be truncated.

[0203] In some embodiments, the viral genome comprises a promoter that results in expression in one or more cells and / or tissues. In some embodiments, the promoter is a ubiquitous promoter. In some embodiments, a promoter that drives or promotes expression in most mammalian tissues includes, but is not limited to. a human elongation factor la-subunit (EFla) promoter, a cytomegalovirus (CMV) immediate-early enhancer and / or promoter, a chicken P-actin (CBA) promoter, a CAG promoter, a CAG derivative promoter, a glucuronidase (GUSB) promoter, and a ubiquitin C (UBC) promoter. A CAG promoter typically comprises: (C) the cytomegalovirus early enhancer element; (A) the promoter, the first exon, and the first intron of the chicken beta-actin gene, and (G) the splice acceptor of the rabbit beta-globin gene. In some embodiments, a derivative of a CAG promoter may comprise (i) a CMVie enhancer and a beta-actin promoter or (ii) a beta-actin promoter and an intron sequence.

[0204] In some embodiments, the viral genome comprises a native GBA1 promoter.

[0205] In some embodiments, the viral genome comprises a nervous system-specific promoter, e.g., a promoter that results in expression of GBA1 in a neuron, an astrocyte, and / or an oligodendrocyte. Nonlimiting examples of tissue-specific expression elements for neurons include synapsin (Syn) or synapsin 1 (Synl), e.g., human synapsin or synapsin 1.

[0206] In some embodiments, the promoter may be less than 1 kb.

[0207] In some embodiments, the promoter may be a combination of tw o or more components of the same or different starting or parental promoters such as, but not limited to, CMV and CBA. Each component may have a length of 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 381. 382, 383, 384, 385, 386, 387, 388, 389, 390, 400, 410, 420, 430, 440, 450,460, 470, 480, 490, 500, 510. 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660,670, 680, 690, 700, 710, 720. 730, 740, 750, 760, 770, 780, 790, 800. or more than 800 nucleotides. Each component may have a length of 200-300, 200-400, 200-500, 200-600, 200-700, 200-800, 300-400, SOO- SOO, 300-600, 300-700, 300-800, 400-500, 400-600. 400-700, 400-800. 500-600, 500-700. 500-800, 600- 700, 600-800 or 700-800 nucleotides. In some embodiments, the promoter is a combination of a 380 nucleotide CMV-enhancer sequence and a 260 nucleotide CBA-promoter sequence.

[0208] In some embodiments, the viral genome comprises an enhancer. In some embodiments, the enhancer comprises a CMVie enhancer.

[0209] In some embodiments, the viral genome comprises a CMVie enhancer and a CBA promoter.In some embodiments, the viral genome comprises a CMVie enhancer and a CMV promoter (e.g., a CMV promoter region). In some embodiments, the viral genome comprises a CMVie enhancer, a CBA promoter or functional variant thereof, and an intron.

[0210] In some embodiments, the viral genome comprises an engineered promoter.

[0211] hi some embodiments, a CBA promoter is used in a viral genome of an AAV particle described herein. In some embodiments, a CBA promoter is used in an AAV particle comprising a viral genome encoding a GBA1 protein.

[0212] In some embodiments, the intron may be 100-600 nucleotides in length. In some embodiments, the intron is 566 nucleotides in length.

[0213] In some embodiments, the AAV vector may comprise a beta-globin intron or a fragment or variant thereof. In some embodiments, the intron comprises one or more human beta-globin sequences (e.g., including fragments / variants thereol). In some embodiments, the promoter may be a CBA promoter. In some embodiments, the promoter comprises a CMV promoter. In some embodiments, the promoter comprises a minimal CBA promoter.

[0214] In some embodiments, the GBAl-encoding sequence may be located downstream of an intron in an expression vector such as a beta globin intron or others known in the art.

[0215] In certain embodiments, the intron sequence is not an enhancer sequence. In some embodiments, the intron sequence is not a sub-component of a promoter sequence. In some embodiments, the intron sequence is a sub-component of a promoter sequence.Viral Genome Component: miR Binding Site-encoding sequence

[0216] Tissue- or cell-specific expression of the AAV viral particles of the disclosure can be enhanced by introducing tissue- or cell-specific regulatory sequences, e.g., promoters, enhancers. microRNA binding sites, e.g., a detargeting site. Without being bound by theory , it is believed that an encoded miR binding site can modulate, e.g., prevent, suppress, or otherwise inhibit, the expression of a gene of interest in the viral genome of the disclosure, based on the expression of the corresponding endogenous microRNA (miRNA) or a corresponding controlled exogenous miRNA in a tissue or cell, e.g., a non-targeting cell or tissue. In some embodiments, a miR binding site modulates, e.g., reduces, expression of GBA1 in a cell or tissue where the corresponding mRNA is expressed. In some embodiments, the miR binding site modulates, e.g., reduces, expression of GBA1 in a cell or tissue of tire DRG, liver, heart, hematopoietic lineage, or a combination thereof.

[0217] In some embodiments, the viral genome of an AAV particle described herein...

Claims

CLAIMSWe claim:

1. An adeno-associated virus (AAV) particle comprising an AAV capsid variant and a viral genome, wherein the viral genome comprises a [3-ghicocerebrosidase 1 (GBAl)-encoding sequence and the AAV capsid variant comprises an amino acid sequence having the formula [N 1]-[N2]-[N3] (SEQ ID NO: 4681) in loop VIII; wherein [N2] comprises the amino acid sequence of DWHR (SEQ ID NO: 4682); and wherein:(i) [Nl] comprises amino acids Xi, X2, X3, and X4, wherein X4is Q. K, E, S, P, R, N, or H; and / or(ii) [N3] comprises amino acids X5, X6, and X?, wherein X5is I, V, T, M, S, N, L, or F.

2. The AAV particle of claim 1, wherein the AAV capsid variant is an AAV9 capsid variant.

3. The AAV particle of claim 1 or claim 2. wherein the AAV capsid variant comprises an amino acid sequence that is at least 95% identical to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

4. The AAV particle of any one of claims 1-3, wherein loop VIII is present at amino acids comprising those corresponding to positions 580-599 of the amino acid sequence of SEQ ID NO: 981.

5. The AAV particle of any one of claims 1-4, wherein:(i) Xi is T, S, R, A, I, C, N. K, L, or Q;(ii) X2is N, T. G, V. S. Y, K.

1. H, D. or F; and(iii) X3is T. N, K, D, I. S, P, A. Y, E. V, L, M, R, H. Q, or C.

6. The AAV particle of any one of claims 1-5, wherein:(i) X6is A. Y, P, N. S, T, G. E, V. W, F, or Q; and(ii) X7is Q, G, N, K, H. R, E, L. P, or M.

7. The AAV particle of any one of claims 1-6, wherein [Nl] comprises the amino acid sequence of TNTQ (SEQ ID NO: 4688).

8. The AAV particle of any one of claims 1-7, wherein [N3] comprises the amino acid sequence of IAQ.

9. The AAV particle of any one of claims 1-8, wherein [N1]-[N2]-[N3] comprises the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343).

10. The AAV particle of any one of claims 1-9, wherein:(i) [Nl] is present at amino acids corresponding to positions 582-585 of the amino acid sequence of SEQ ID NO: 981;(ii) [N2] is present at amino acids corresponding to positions 586-589 of the amino acid sequence of SEQ ID NO: 981: and(iii) [N3] is present at amino acids corresponding to positions 590-592 of the amino acid sequence of SEQ ID NO: 981.

11. The AAV particle of any one of claims 1-10, wherein the AAV capsid variant further comprises [N4] comprising amino acids X8. X9. Xio, and Xu, wherein:(i) X8is T, S. N, P, A. or I;(ii) X9is G, N, D. R, V. A, S, or Q;(iii) Xio is W. S, C, R. L, or G; and / or(iv) Xu is V. A, S, I, C. G, D. F, L, or T.

12. The AAV particle of claim 11, wherein [N4] comprises the amino acid sequence of TGWV (SEQ ID NO: 5066).

13. The AAV particle of claim 11 or claim 12, wherein [N4] is present at amino acids corresponding to positions 593-596 of the amino acid sequence of SEQ ID NO: 981.

14. The AAV particle of any one of claims 11-13, wherein [N1]-[N2]-[N3]-[N4] comprises the amino acid sequence of TNTQDWHRIAQTGWV (SEQ ID NO: 201).

15. An adeno-associated virus (AAV) particle comprising:(i) a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence: and(ii) an AAV capsid variant comprising an amino acid sequence that is at least 95% identical to positions 203-736 of the amino acid sequence of SEQ ID NO: 981, wherein the AAV capsid variant comprises:T at an amino acid corresponding to position 584 of the amino acid sequence of SEQ ID NO: 981;D at an amino acid corresponding to position 586 of the amino acid sequence of SEQ ID NO:W at an amino acid corresponding to position 587 of the amino acid sequence of SEQ ID NO:981;H at an amino acid corresponding to position 588 of the amino acid sequence of SEQ ID NO: 981;R at an amino acid corresponding to position 589 of the amino acid sequence of SEQ ID NO: 981; andI at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 981.

16. The AAV particle of any one of claims 1-15, wherein the AAV capsid variant comprises an amino acid sequence that is at least 95% identical to positions 138-736 of the amino acid sequence of SEQ ID NO: 981.

17. An adeno-associated virus (AAV) particle comprising:(i) a viral genome comprising a P-glucocerebrosidase 1 (GBAl)-encoding sequence; and(ii) an AAV capsid variant comprising an amino acid sequence that is at least 95% identical to positions 138-736 of the amino acid sequence of SEQ ID NO: 981, wherein the AAV capsid variant comprises:T at an amino acid corresponding to position 584 of the amino acid sequence of SEQ IDNO: 981;D at an amino acid corresponding to position 586 of the amino acid sequence of SEQ ID NO: 981;W at an amino acid corresponding to position 587 of the amino acid sequence of SEQ ID NO: 981;H at an amino acid corresponding to position 588 of the amino acid sequence of SEQ ID NO: 981;R at an amino acid corresponding to position 589 of the amino acid sequence of SEQ ID NO: 981; andI at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 981.

18. The AAV particle of any one of claims 1-17, wherein the AAV capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 981.

19. An adeno-associated virus (AAV) particle comprising:(i) a viral genome comprising a p-glucocerebrosidase 1 (GBAl)-encoding sequence; and(ii) an AAV capsid variant comprising an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

981. wherein the AAV capsid variant comprises:T at an amino acid corresponding to position 584 of the amino acid sequence of SEQ IDNO: 981;D at an amino acid corresponding to position 586 of the amino acid sequence of SEQ ID NO: 981;W at an amino acid corresponding to position 587 of the amino acid sequence of SEQ ID NO: 981;H at an amino acid corresponding to position 588 of the amino acid sequence of SEQ ID NO: 981;R at an amino acid corresponding to position 589 of the amino acid sequence of SEQ ID NO: 981; andI at an amino acid corresponding to position 590 of the amino acid sequence of SEQ ID NO: 981.

20. The adeno-associated virus (AAV) particle of claim 19, wherein the AAV capsid variant comprises:(i) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 981;(ii) an amino acid sequence that is at least 99% identical to positions 138-736 of the amino acid sequence of SEQ ID NO: 981; and / or(iii) an amino acid sequence that is at least 99% identical to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

21. The AAV particle of any one of claims 15-20, wherein tire AAV capsid variant comprises the amino acid sequence of TQDWHRI (SEQ ID NO: 941).

22. The AAV particle of any one of claims 15-21, wherein tire AAV capsid variant comprises at least 8, at least 9, or at least 10 consecutive amino acids from the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343).

23. The AAV particle of any one of claims 15-22, wherein the AAV capsid variant comprises the amino acid sequence of TNTQDWHRIAQ (SEQ ID NO: 343) present at amino acids corresponding to positions 582-592 of the amino acid sequence of SEQ ID NO: 981.

24. The AAV particle of any one of claims 21-23. wherein the amino acid sequence of TQDWHRI (SEQ ID NO: 941) or TNTQDWHRIAQ (SEQ ID NO: 343) is present in loop VIII. wherein loop VIII comprises amino acids 580-599 of the amino acid sequence of SEQ ID NO: 981.

25. The AAV particle of any one of claims 1-24, wherein the AAV capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 981;(ii) the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981; and / or(iii) the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

26. The AAV particle of any one of claims 1-25, wherein the viral genome encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1775. 1740. 1742. 1744, 1746, or 1748 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

27. The AAV particle of any one of claims 1-26, wherein the viral genome encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1775 or an amino acid sequence that is at least 90% identical (e.g.. at least 90%, at least 91%. at least 92%. at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

28. The AAV particle of any one of claims 1-27. wherein the viral genome encodes a wildtype GBA1 protein.

29. The AAV particle of any one of claims 1-28, wherein the viral genome encodes a human GBA1 protein, a dog GBA1 protein, an equine GBA1 protein, or a monkey GBA1 protein.

30. The AAV particle of any one of claims 1-29, wherein the viral genome encodes a wildtype human GBA1 protein.

31. The AAV particle of any one of claims 1-30, wherein the viral genome docs not encode a hemagglutinin (HA) tag.

32. The AAV particle of any one of claims 1-31, wherein the GBAl-encoding sequence comprises SEQ ID NO: 2002 or SEQ ID NO: 1773, or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

33. The AAV particle of any one of claims 1-32, where the viral genome further comprises a signal sequence-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 2005 or SEQ ID NO: 1850. or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

34. The AAV particle of any one of claims 1-33, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 1772, or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

35. The AAV particle of any one of claims 1-34, wherein the viral genome encodes a GBA1 protein comprising the amino acid sequence of SEQ ID NO: 1774 or an amino acid sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

36. The AAV particle of any one of claims 1-35, wherein the viral genome comprises a promoter operably linked to the GBA1 -encoding sequence.

37. The AAV particle of claim 36, wherein the promoter comprises a human elongation factor 1 alpha (EF-la) promoter, a chicken [Lactin (CBA) promoter, a CAG promoter, a CAG derivative promoter, a CMV immediate-early enhancer and / or promoter, a CMV promoter, a [3 glucuronidase (GUSB) promoter, a ubiquitin C (UBC) promoter, a neuron-specific enolase (NSE) promoter, a platelet- derived growth factor (PDGF) promoter, a platelet-derived growth factor B-chain (PDGF-0) promoter, an intercellular adhesion molecule 2 (ICAM-2) promoter, a synapsin (Syn) promoter, a methyl-CpG binding protein 2 (MeCP2) promoter, a Ca2+ / calmodulin-dependent protein kinase II (CaMKII) promoter, a metabotropic glutamate receptor 2 (mGluR2) promoter, a neurofilament light chain (NFL) promoter, aneurofilament heavy chain (NFH) promoter, a -globin minigene n 2 promoter, a preproenkephalin (PPE) promoter, an enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2) promoter, a glial fibrillary acidic protein (GFAP) promoter, a myelin basic protein (MBP) promoter, a cardiovascular promoter (e.g., aMHC, cTnT, and CMV-MLC2k), a liver promoter (e.g., hAAT, TBG), a skeletal muscle promoter (e.g., desmin, MCK, C512), or a fragment, e.g., a truncation, or a functional variant thereof.

38. The AAV particle of claim 36 or claim 37, wherein the promoter comprises a CBA promoter.

39. The AAV particle of any one of claims 36-38, wherein the promoter comprises the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

40. The AAV particle of any one of claims 1-39, wherein the viral genome further comprises an enhancer.

41. The AAV particle of claim 40, wherein the enhancer comprises a CMV immediate-early (CMVie) enhancer; optionally wherein the enhancer comprises the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

42. The AAV particle of any one of claims 1-41, wherein the viral genome further comprises an intron; optionally wherein the intron comprises the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%. at least 92%, at least 93%, at least 94%. at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto.

43. The AAV particle of any one of claims 1-42. wherein the viral genome further comprises a polyadenylation (poly A) region; optionally wherein the polyA region comprises the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

44. The AAV particle of any one of claims 1-43, wherein the viral genome further comprises an in verted terminal repeat (ITR); optionally wherein the ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or SEQ ID NO: 1830 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

45. The AAV particle of claim 44, wherein the viral genome comprises a 5’ ITR and a 3’ ITR, wherein the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%. at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto, and the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%. at least 93%, at least 94%. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

46. The AAV particle of any one of claims 1-45, wherein the viral genome further comprises a nucleotide sequence encoding one or more miR183 binding sites.

47. The AAV particle of any one of claims 1-46, wherein the viral genome encodes at least four miR183 binding sites.

48. The AAV particle of claim 47, wherein each of the at least four miR183 binding sites is separated by a spacer.

49. The AAV particle of any one of claims 45-48. wherein each of the miR183 binding sites is encoded by a nucleotide sequence that comprises the nucleotide sequence of SEQ ID NO: 1847 or a nucleotide sequence that has up to three modifications relative thereto.

50. The AAV particle of any one of claims 1-45, wherein the viral genome further comprises a nucleotide sequence encoding a miR183 binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto.

51. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; and(iv) a 3’ ITR.

52. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; and(v) a 3’ ITR.

53. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) an intron;(v) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto; and(vi) a 3’ ITR.

54. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vi) a 3‘ ITR.

55. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence drat is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a poly adenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vii) a 3’ ITR.

56. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5' inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%. at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence tiiat is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a poly adenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto.

57. The AAV particle of any one of claims 51-56. wherein:(i) the 5' ITR comprises the nucleotide sequence of SEQ ID NO: 1829;(ii) the enhancer comprises the nucleotide sequence of SEQ ID NO: 1831;(iii) the promoter comprises the nucleotide sequence of SEQ ID NO: 1834;(iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842;(v) the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001;(vi) the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846; and (viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

58. The AAV particle of any one of claims 1-56, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

59. The AAV particle of any one of claims 1-45 or 51-58, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006.

60. The AAV particle of any one of claims 1-45 or 51-59, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2006.

61. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto;(iv) a nucleotide sequence encoding at least one miR183 binding site; and(v) a 3’ ITR.

62. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) a promoter;(iii) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(iv) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and(v) a 3’ ITR.

63. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(v) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and(vi) a 3’ ITR.

64. The AAV particle of any one of claims 1-25. wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer;(iii) a promoter;(iv) an intron;(v) the GB Al -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and(vii) a 3’ ITR.

65. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence drat is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a nucleotide sequence encoding a miR183 binding site series, wherein the miR183 binding site series comprises at least one miR183 binding site and at least one spacer sequence; and(vii) a 3’ ITR.

66. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR);(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%. at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBAl-encoding sequence, wherein the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a nucleotide sequence encoding a miR183 binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and(vii) a 3’ ITR.

67. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto;(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. or at least 99% identical) thereto;(vi) a nucleotide sequence encoding at least one miR183 binding site, wherein the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of SEQ ID NO: 1847;(vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g.. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

68. The AAV particle of any one of claims 1-25, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 1829 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(ii) an enhancer comprising the nucleotide sequence of SEQ ID NO: 1831 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) a promoter comprising the nucleotide sequence of SEQ ID NO: 1834 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto;(iv) an intron comprising the nucleotide sequence of SEQ ID NO: 1842 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto;(v) the GBA1 -encoding sequence, wherein the GBA1 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001 or SEQ ID NO: 2002 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vi) a nucleotide sequence encoding a miR183 binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(vii) a polyadenylation (poly A) region comprising the nucleotide sequence of SEQ ID NO: 1846 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%. at least 97%, at least 98%, or at least 99% identical) thereto; and(viii) a 3’ ITR comprising the nucleotide sequence of SEQ ID NO: 1830 or a nucleotide sequence that is at least 95% identical (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

69. The AAV particle of claim 68, wherein:(i) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 1829;(ii) the enhancer comprises the nucleotide sequence of SEQ ID NO: 1831 ;(iii) the promoter comprises the nucleotide sequence of SEQ ID NO: 1834;(iv) the intron comprises the nucleotide sequence of SEQ ID NO: 1842;(v) the GBAl-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 2001;(vi) the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 1849;(vii) the poly A region comprises the nucleotide sequence of SEQ ID NO: 1846; and(viii) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 1830.

70. The AAV particle of any one of claims 1-68, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

71. The AAV particle of claim 70, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2007.

72. The AAV particle of claim 71, wherein the viral genome consists of the nucleotide sequence of SEQ ID NO: 2007.

73. A cell comprising the AAV particle of any one of claims 1-72, optionally wherein the cell is a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

74. A method of making the AAV particle of any one of claims 1-72, the method comprising:(i) providing a cell comprising the viral genome comprising a GBAl-encoding sequence and a nucleic acid encoding the AAV capsid variant; and(ii) incubating the cell under conditions suitable to encapsulate the viral genome in the AAV capsid variant; thereby making the AAV particle.

75. The method of claim 74, wherein the viral genome comprises:(i) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g.. at least 90%, at least 91%. at least 92%. at least 93%, at least 94%. at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(ii) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(iii) the nucleotide sequence of SEQ ID NO: 1812 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; or(iv) the nucleotide sequence of SEQ ID NO: 1828 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises:(a) the amino acid sequence of SEQ ID NO: 981 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(b) the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981 or an amino acid sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto: and / or(c) the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981 or an amino acid sequence that is at least 90% identical (e.g.. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto.

76. The method of claim 74. wherein the viral genome comprises:(i) the nucleotide sequence of SEQ ID NO: 2006 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; or(ii) the nucleotide sequence of SEQ ID NO: 2007 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981, the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO: 981, and / or the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

77. The method of claim 74, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 2006 or SEQ ID NO: 2007 and the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981, the amino acid sequence according to positions 138-736 of the amino acid sequence of SEQ ID NO:

981. and / or the amino acid sequence according to positions 203-736 of the amino acid sequence of SEQ ID NO: 981.

78. The method of any one of claims 74-77, further comprising, prior to step (i). introducing a nucleic acid molecule comprising the viral genome into the cell.

79. The method of any one of claims 74-78, further comprising, prior to step (i). introducing the nucleic acid encoding the AAV capsid variant into the cell.

80. The method of any one of claims 74-79, wherein the cell comprises a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an SI9 cell), or a bacterial cell.

81. A pharmaceutical composition comprising the AAV particle of any one of claims 1-72 and a pharmaceutically acceptable excipient.

82. A method of delivering an AAV particle encoding a GBA1 protein to a cell, comprising administering an effective amount of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72.

83. The method of claim 82, wherein the cell is in a subject, optionally where in subject has. has been diagnosed with having, or is at risk of having a GBA1 -related disorder.

84. A method of treating a GBA1 -related disorder in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72.

85. The method of claim 84, wherein the subject has, has been diagnosed with having, or is at risk of having the GB Al -related disorder.

86. The method of any one of claims 83-85. wherein the GBAl-related disorder is a GBAl-related neurodegenerative or neuromuscular disorder.

87. The method of any one of claims 83-86. wherein the GBAl-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g.. GD type 1, GD type 2. or GD type 3), Dementia with Lewy Bodies (DLB). Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome.

88. The method of any one of claims 83-87, wherein the GBAl-related disorder is PD.

89. The method of any one of claims 83-87, wherein the GBAl-related disorder is LBD.

90. The method of any one of claims 83-87, wherein the GBAl-related disorder is DLB.

91. The method of any one of claims 83-87, wherein the GBAl-related disorder is GD.

92. A method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Parkinson’s Disease (PD), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72.

93. The method of claim 92, wherein the subject has, has been diagnosed with having, or is at risk of having PD.

94. A method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Lewy Body Dementia (LBD), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72.

95. The method of claim 94, wherein the subject has, has been diagnosed with having, or is at risk of having LBD.

96. A method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Dementia with Lewy Bodies (DLB), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72.

97. The method of claim 96, wherein the subject has, has been diagnosed with having, or is at risk of having DLB.

98. A method of treating a GBAl-related disorder in a subject, wherein the GBAl-related disorder is Gaucher Disease (GD), comprising administering to the subject an effective amount of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72.

99. The method of claim 98, wherein the subject has. has been diagnosed with having, or is at risk of having GD.

100. The method of claim 98 or claim 99, wherein the GD is GD type 1.

101. The method of claim 98 or claim 99, wherein the GD is GD type 2.

102. The method of claim 98 or claim 99, wherein the GD is GD type 3.

103. The method of any one of claims 83-102, wherein the subject has one or more mutations in theGBA1 gene.

104. The method of any one of claims 83-103, wherein the subject has lower GCase activity prior to administration of the pharmaceutical composition or AAV particle as compared to GCase activity in an individual who does not have a GBAl-related disorder, optionally wherein the level of GCase activity is measured by a 4-MUG assay or a SensoLyte Blue Glucocerebrosidase assay.

105. The method of any one of claims 84-104, wherein the treating results in prevention of progression of the GBAl-related disorder in the subject.

106. The method of any one of claims 84-105, wherein the treating results in amelioration of at least one symptom of the GBAl-related disorder in the subject.

107. The method of claim 106, wherein the at least one symptom comprises developmental delay, progressive encephalopathy, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, generalized weakness, trembling of a limb, depression, visual hallucinations, cognitive decline, or a combination thereof.

108. The method of any one of claims 105-107, wherein the treating results in a change in one or more biomarkers comprising a GCase activity, a level of glucocerebroside and other glycolipids, (e.g., within immune cells such as macrophages), a level of synuclein aggregates (e.g., Lewy bodies), or a combination thereof.

109. The method of any one of claims 83-108, wherein the subject is a human.

110. The method of any one of claims 83-109, wherein the AAV particle or the pharmaceutical composition is delivered to a cell or tissue of the central nervous system (CNS) in the subject.

111. The method of claim 110. wherein the cell or tissue of the CNS is a cell or tissue of the amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region).

112. The method of any one of claims 82-108, wherein the AAV particle or the pharmaceutical composition is delivered to a peripheral cell or tissue in the subject.

113. The method of claim 112, wherein the peripheral cell or tissue is a cell or tissue of the heart, skeletal muscle, sympathetic ganglia, and / or plasma.

114. The method of any one of claims 83-113, wherein the AAV particle or pharmaceutical composition is delivered to the subject via intravenous administration.

115. The method of any one of claims 83-114, further comprising evaluating, e.g., measuring, the level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) in the subject, e.g.. in a cell, tissue, or fluid of the subject.

116. The method of claim 115, wherein the level of GBA1 protein expression is measured by an enzyme-linked immunosorbent assay (ELISA), a Western blot, or an immunohistochemistry assay.

117. The method of claim 115 or claim 116, wherein evaluating the subject’s level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) is performed prior to and / or subsequent to administration of the pharmaceutical composition or AAV particle, optionally wherein the subject’s level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) prior to administration is compared to the subject’s level of GBA1 expression (e.g., GBA1 gene expression, GBA1 mRNA expression, and / or GBA1 protein expression) subsequent to administration.

118. The method of any one of claims 115-117, comprising evaluating the level of GBA1 expression in a cell or tissue of the central nervous system.

119. The method of any one of claims 115-118. wherein the subject’s level of GBA1 protein expression subsequent to administration is increased relative to the subject’s level of GBA1 protein expression prior to administration.

120. The method of any one of claims 83-119, further comprising evaluating, e.g., measuring, the level of GCase activity in the subject.

121. The method of any one of claims 83-120, wherein administering the pharmaceutical composition or AAV particle to the subject results in an increase in:(i) GCase activity in a cell, tissue, (e.g., a cell or tissue of the CNS, e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary' complex, nucleus ambiguus. oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g.. cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)), and / or fluid (e.g., CSF and / or serum) of the subject relative to baseline and / or relative to GCase activity in a cell, tissue, or fluid of an individual with a GBA1 -related disorder who has not been administered the pharmaceutical composition or AAV particle;(ii) the number and / or level of viral genomes (VG) per cell in a CNS tissue (e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g.. Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus,hippocampus, inferior colliculus, inferior olivary' complex, nucleus ambiguus, oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g., cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)) of the subject relative to the number and / or level of VG per cell in a peripheral tissue of the subject; and / or(iii) GBA1 mRNA expression in a cell or tissue (e g., a cell or tissue of the CNS, e.g., amygdala, brainstem, caudate, central grey, cerebellum (e.g., Purkinje cell layer and deep cerebellar nuclei), cortex (e.g., frontal cortex, motor cortex, perirhinal cortex, sensory cortex, temporal cortex, visual cortex), external cuneate nucleus, geniculate nucleus, globus pallidus, gracile nucleus, hilus of the dentate gyrus, hippocampus, inferior colliculus, inferior olivary complex, nucleus ambiguus. oculomotor nucleus, putamen, substantia nigra, thalamus, ventral palladium, vestibular nucleus, and / or spinal cord (e.g.. cervical spinal cord region, lumbar spinal cord region, or thoracic spinal cord region)) of the subject relative to baseline and / or relative to GBA1 mRNA expression in a cell or tissue of an individual with a GBA1 -related disorder who has not been administered the pharmaceutical composition or AAV particle.

122. The method of any one of claims 83-121, further comprising administering to the subject at least one additional agent and / or therapy.

123. The method of claim 122. wherein the at least one additional agent and / or therapy comprises an agent and / or therapy suitable for treating a GB Al -related disorder.

124. The method of claim 123, wherein the GB Al -related disorder is PD, LBD, GD, or DLB, wherein, optionally, the GD is GD type 1, GD type 2, or GD type 3.

125. The method of any one of claims 122-124, wherein the at least one additional agent and / or dicrapy is suitable for treating Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e g., GD type 1, GD type 2, or GD type 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD), Multiple System Atrophy (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation ty pe1 (NBIA 1), or Hallervorden-Spatz Syndrome.

126. The method of any one of claims 122-125, wherein the at least one additional agent and / or therapy comprises enzyme replacement therapy (ERT) (e.g., imiglucerase, velaglucerase alfa, or taliglucerase alfa); substrate reduction therapy (SRT) (e.g.. eliglustat or miglustat), levodopa, carbidopa, Safinamide. a dopamine agonist (e.g.. pramipexole, rotigotine, or ropinirole), a dopamine antagonist (e.g., quetiapine, clozapine), an anticholinergic (e.g., benztropine or trihexyphenidyl), a cholinesterase inhibitor (e.g.. rivastigmine, donepezil, or galantamine), an N-methyl-d-aspartate (NMDA) receptor antagonist (e.g.. memantine), or a combination thereof.

127. The method of any one of claims 83-126, further comprising administering an immunosuppressant to the subject.

128. The method of claim 127, wherein the immunosuppressant comprises a corticosteroid (e g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxychloroquine.

129. The method of any one of claims 83-128, further comprising administering a blood transfusion to the subject.

130. The pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72 for use in a method of treating a disorder according to any one of claims 84-129.

131. The pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1-72 for use in the treatment of a GBA1 -related disorder in a subject.

132. The pharmaceutical composition or AAV particle of claim 131. wherein the GBAl-related disorder is Parkinson’s Disease (PD), Parkinson’s Disease Dementia (PDD), Gaucher Disease (GD) (e.g.. GD t pe 1, GD type 2, or GD type 3), Dementia with Lewy Bodies (DLB), Lewy Body Dementia (LBD). Multiple System Atrophy (MSA). Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden-Spatz Syndrome.

133. The pharmaceutical composition or AAV particle of claim 131 or claim 132, wherein the GBAl- related disorder is PD.

134. The pharmaceutical composition or AAV particle of claim 133, wherein the subject has, has been diagnosed with having, or is at risk of having PD.

135. The pharmaceutical composition or AAV particle of claim 131 or claim 132, wherein the GBAl- related disorder is LBD.

136. The pharmaceutical composition or AAV particle of claim 135, wherein the subject has, has been diagnosed with having, or is at risk of having LBD.

137. The pharmaceutical composition or AAV particle of claim 131 or claim 132, wherein the GBA1- related disorder is DLB.

138. The pharmaceutical composition or AAV particle of claim 137, wherein the subject has, has been diagnosed with having, or is at risk of having DLB.

139. The pharmaceutical composition or AAV particle of claim 131 or claim 132, wherein the GBA1- related disorder is GD (e.g., GD type 1, GD type 2, or GD type 3).

140. The pharmaceutical composition or AAV particle of claim 139, wherein the subject has, has been diagnosed with having, or is at risk of having GD (e.g., GD type 1. GD type 2, or GD type 3).

141. Use of the pharmaceutical composition of claim 81 or the AAV particle of any one of claims 1- 72 in the manufacture of a medicament for the treatment of a GBAl-related disorder in a subject.

142. The use of claim 141. wherein the GBAl-related disorder is Parkinson’s Disease (PD). Parkinson’s Disease Dementia (PDD). Gaucher Disease (GD) (e.g.. GD type 1, GD type 2. or GD type 3), Dementia with Lewy Bodies (DLB), Lewy’ Body’ Dementia (LBD), Multiple System Atrophy’ (MSA), Alzheimer’s Disease (AD), Amyotrophic Lateral Sclerosis (ALS), Pure Autonomic Failure, Neurodegeneration with brain iron accumulation type 1 (NBIA 1), or Hallervorden- Spatz Syndrome.

143. The use of claim 141 or claim 142, wherein the GBAl-related disorder is PD.

144. The use of claim 143, wherein the subject has, has been diagnosed with having, or is at risk of having PD.

145. The use of claim 141 or claim 142, wherein the GBAl-related disorder is GD.

146. The use of claim 145, wherein the subject has, has been diagnosed with having, or is at risk of having GD.

147. The use of claim 142, 145, or 146, wherein the GD is GD type 1.

148. The use of claim 142, 145, or 146, wherein the GD is GD type 2.

149. The use of claim 142, 145, or 146, wherein the GD is GD type 3.

150. The use of claim 141 or claim 142, wherein the GBAl-related disorder is LBD.

151. The use of claim 150, wherein the subject has, has been diagnosed with having, or is at risk of having LBD.

152. The use of claim 141 or claim 142, wherein the GBAl-related disorder is DLB.

153. The use of claim 152, wherein the subject has. has been diagnosed with having, or is at risk of having DLB.

Images