Compositions and methods for delivering cyclin-dependent kinase-like 5 protein

Abstract

The disclosure relates to adeno-associated (AAV) particles comprising viral genomes encoding cyclin-dependent kinase-like 5 (CDKL5) proteins and peptides, compositions comprising said AAV particles, and methods for making and delivering said AAV particles to a cell or subject. The AAV particles, compositions, and methods of the present disclosure are useful for the treatment of subjects who have, have been diagnosed with having, or are at risk of having a CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, atypical Rett syndrome, and / or other CDKL5-related disorders or at least one symptom thereof.

Description

Attorney Docket No. 14640.0304-00304COMPOSITIONS AND METHODS FOR DELIVERING CYCLIN-DEPENDENT KINASE-LIKE 5 PROTEINRelated Applications

[0001] This application claims the benefit of and priority to U. S. Provisional Application No. 63 / 714,108, filed on October 30, 2024, 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_0304-00304_SL.xml, was created on October 3, 2025, and is 137,503 bytes in size. The information in electronic format of the Sequence Listing is incorporated herein by reference in its entirety.Field

[0003] Described herein are adeno-associated virus (AAV) particles comprising a viral genome (e.g., recombinant viral genome) encoding cyclin-dependent kinase-like 5 (CDKL5) proteins and peptides. Also described herein are compositions comprising said AAV particles, methods of making said AAV particles, and / or methods of delivering said AAV particles to a cell or subject. In some embodiments, the disclosure provides methods or uses for the treatment of CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome and / or other CDKL5-related disorders.Background

[0004] Cyclin-dependent kinase-like 5 (CDKL5) is a serine / threonine protein kinase and is also known as serine / threonine kinase 9 (STK9). Other aliases for CDKL5 include EIEE2, ISSX, CFAP247, and DEE2. It is encoded by the gene CDKL5 (Ensembl Gene ID No.ENSG00000008086), which is located on the X chromosome.

[0005] The CDKL5 protein is an enzyme and is thought to play an important role in brain development and regulation of response to oxidative stress. CDKL5 is thought to be expressed throughout the cell, including in the nucleus and cytoplasm of soma and dendrites.

[0006] CDKL5 is responsible for phosphorylation of a number of targets. CDKL5 may target and phosphorylate the gene MECP2, which has been characterized as important in theAttorney Docket No. 14640.0304-00304function of neurons and other brain cells, and in the maintenance of neuronal synapses, and is thought to be the causative agent for Rett syndrome. CDKL5 may also target and phosphorylate CEP131, which is thought to play a role in cell proliferation. CDKL5 may also target and phosphorylate MAP IS, DLG5, EB2, and / or ARHGEF, which are microtubule associated proteins, thought to play roles in neuronal division, differentiation, migration, and neurite growth. CDKL5 may also target and phosphorylate AKT and mTOR, which are thought to play roles in cell proliferation, migration and development.

[0007] Without being bound by theory, CDKL5 may be involved in the formation, growth, and migration of neurons. It may also play a role in cell division and / or transmission of chemical signals at neuronal synapses.

[0008] Mutations in CDKL5 are known to cause disease in subjects, e.g., human subjects. CDKL5 mutations lead to CDKL5 deficiency disorder (CDD). CDD is a neurodevelopmental disorder. It is characterized by nervous system symptoms including epilepsy (e.g., early-onset epilepsy), autism, deficits in cognition, limited motor skills, sleep difficulties and / or visual impairment. It is also characterized by low muscle tone and gastrointestinal reflux.

[0009] CDD can manifest with a broad array of clinical manifestations. Clinical manifestations of CDD comprise behavioral symptoms such as episodes of laughing or crying that occur for what appears to be no reason, hypersensitivity to touch, and disrupted sleep. Clinical manifestations also comprise facial appearance changes including microcephaly; a high, broad forehead; large, deep-set eyes; smaller-than normal space between the nose and upper lip; an upturned nose; fuller lips; and / or widely-spaced teeth. Further clinical manifestations include difficulties standing and walking, small, cold feet, lack of or poor eye contact, frequent sideways glances, and cortical visual impairment or cortical blindness. Other manifestations include bruxism, limited or absent speech, difficulties eating, stereotypies, limited ability to make small, focused hand movements, gastroesophageal reflux and constipation.

[0010] CDD has an incidence of 1 in 42000 births in the USA, and 85% of cases occur in females. Typically, CDD patients are fully reliant on caregivers for the duration of their lives.

[0011] CDD is typically caused by de novo mutations in CDKL5. CDKL5 mutations in CDD patients result in a reduced amount of functional CDKL5 protein.

[0012] Existing treatments for patients with CDD focus on alleviating symptoms such as seizures. To date, there are limited treatments available for patients with CDD, and deliveryAttorney Docket No. 14640.0304-00304of treatments to the central nervous system (CNS) remains a significant challenge in the development of new and effective therapies.

[0013] The current standard of care for CDD is first line treatment with an anti-epileptic drug. Anti-epileptic drugs are known in the art, and include valproate and levetiracetam. Second line treatment for refractory patients comprises first line treatment plus additional anti-epileptic drugs such as clobazam and / or lamotrigine in combination with ganaxolone. However, ganaxolone has been reported to have short durability in treating subjects with CDD. Third line treatment introduces additional anti-epileptic drugs, such as topiramate, and / or alternative interventions, such as ketogenic diet and / or vagal nerve stimulation.However, seizures in CDD are commonly refractory to known treatments.

[0014] Therefore, a need remains for improved therapies and treatments that target the cause of CDD. In particular, there remains a need for pharmaceutical compositions and methods to treat CDD that can be delivered to the CNS for the treatment of CDD, and to ameliorate deficiencies of CDKL5 in subjects, e.g., human subjects.

[0015] Prior attempts at providing AAV capsids with improved properties, e.g., improved tropism suitable for delivery to the brain or CNS, have met with limited success. As such, there remains a need for effective methods of treatment using AAV capsid variants that are capable of delivering a payload of interest, e.g., CDKL5, to a target cell or tissue, e.g., a CNS cell or tissue.Summary

[0016] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25 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 thereto; and wherein the AAV9 capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 5) present at amino acids corresponding to positions 254-259 of the amino acid sequence of SEQ ID NO: 25, the amino acid E at position 249 of the amino acid sequence of SEQ ID NO: 25, and the amino acid V at position 251 of the amino acid sequence of SEQ ID NO: 25.

[0017] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genomeAttorney Docket No. 14640.0304-00304comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25 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 thereto; and wherein the AAV9 capsid variant comprises the amino acid sequence of ENVSGSPHSKA (SEQ ID NO: 8) present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25, optionally wherein the AAV9 capsid variant comprises the amino acid sequence of KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25.

[0018] In some embodiments, the AAV9 capsid variant comprises: (i) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 1, wherein the amino acid sequence comprises ENVSGSPHSKA (SEQ ID NO: 8) present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1, optionally wherein the amino acid sequence comprises KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1; and / or (ii) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 24, wherein the amino acid sequence comprises ENVSGSPHSKA (SEQ ID NO: 8) present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24, optionally wherein the amino acid sequence comprises KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24.

[0019] In some embodiments, the AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 1; (ii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 24; and / or (iii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 25.Attorney Docket No. 14640.0304-00304

[0020] In some embodiments, the AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 1; (ii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 24; and / or (iii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 25. In some embodiments, the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 1; (ii) the amino acid sequence of SEQ ID NO: 24; and / or (iii) the amino acid sequence of SEQ ID NO: 25.

[0021] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27 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 thereto; and wherein the AAV9 capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 5) present at amino acids corresponding to positions 254-259 of the amino acid sequence of SEQ ID NO: 27, the amino acid E at position 249 of the amino acid sequence of SEQ ID NO: 27, the amino acid R at position 250 numbered according to the amino acid sequence of SEQ ID NO: 27, and the amino acid V at position 251 of the amino acid sequence of SEQ ID NO: 27.

[0022] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27 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 thereto; and wherein the AAV9 capsid variant comprises the amino acid sequence of ERVSGSPHSKA (SEQ ID NO: 10) present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27, optionally wherein the AAV9 capsid variant comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27.

[0023] In some embodiments, the AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 90% (e.g., at least 90%, at least 91%, at least 92%, at least 93%, atAttorney Docket No. 14640.0304-00304least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 4, wherein the amino acid sequence comprises ERVSGSPHSKA (SEQ ID NO: 10) present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4, optionally wherein the amino acid sequence comprises KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4; and / or (ii) an amino acid sequence that is 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 26, wherein the amino acid sequence comprises ERVSGSPHSKA (SEQ ID NO: 10) present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26, optionally wherein the amino acid sequence comprises KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26.

[0024] In some embodiments, AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 95% (at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 4; (ii) an amino acid sequence that is at least 95% (at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 26; and / or (iii) an amino acid sequence that is at least 95% (at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 27. In some embodiments, the AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 99% identical to SEQ ID NO: 4; (ii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 26; and / or (iii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 27. In some embodiments, the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 4; (ii) the amino acid sequence of SEQ ID NO: 26; and / or (iii) the amino acid sequence of SEQ ID NO: 27.

[0025] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47 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 thereto; andAttorney Docket No. 14640.0304-00304wherein the AAV9 capsid variant comprises HDSPHK (SEQ ID NO: 12) present at amino acids 252-257 of the amino acid sequence of SEQ ID NO: 47 and comprises the amino acid R at position 388, the amino acid T at position 390, the amino acid L at position 392, the amino acid Q at position 393, and the amino acid L at position 394, each numbered according to the amino acid sequence of SEQ ID NO: 47.

[0026] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE); wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47 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 thereto; and wherein the AAV9 capsid variant comprises HDSPHK (SEQ ID NO: 12) present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47, optionally wherein the AAV9 capsid variant comprises the amino acid sequence of KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47.

[0027] In some embodiments, the AAV9 capsid variant comprises: (i) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 45, wherein the amino acid sequence comprises HDSPHK (SEQ ID NO: 12) present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45, optionally wherein the amino acid sequence comprises KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45; and / or (ii) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 46, wherein the amino acid sequence comprises HDSPHK (SEQ ID NO: 12) present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 and comprises the aminoAttorney Docket No. 14640.0304-00304acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46, optionally wherein the amino acid sequence comprises KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46.

[0028] In some embodiments, the AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 45; (ii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 46; and / or (iii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 47. In some embodiments, the AAV9 capsid variant comprises: (i) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 45; (ii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 46; and / or (iii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 47. In some embodiments, the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 45; (ii) the amino acid sequence of SEQ ID NO: 46; and / or (iii) the amino acid sequence of SEQ ID NO: 47.

[0029] In some embodiments, the recombinant viral genome encodes a wildtype CDKL5 protein. In some embodiments, the recombinant viral genome encodes a human CDKL5 protein.

[0030] In some embodiments, the encoded CDKL5 comprises the amino acid sequence of SEQ ID NO: 14 or any one of SEQ ID NOs: 34-37.

[0031] In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide sequence that is at least 70% (e.g., at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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 CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identicalAttorney Docket No. 14640.0304-00304thereto. In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15.

[0032] In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16 or a nucleotide sequence that is at least 98% (e.g., at least 98% or at least 99%) identical thereto. In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16.

[0033] In some embodiments, the WPRE is positioned 3’ relative to the CDKL5-encoding sequence. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or any one of SEQ ID NOs: 58-62, or a nucleotide 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 thereto. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 19.

[0034] In some embodiments, the recombinant viral genome further comprises a promoter operably linked to the CDKL5-encoding sequence. In some embodiments, the promoter is a human synapsin 1 (hSYNl) promoter or a chicken P-actin hybrid (CBh) promoter. In some embodiments, the promoter is a hSYNl promoter. In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto. In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 18.

[0035] In some embodiments, the recombinant viral genome further comprises a polyadenylation (poly A) sequence. In some embodiments, the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto. In some embodiments, the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20.

[0036] In some embodiments, the recombinant viral genome further comprises at least one inverted terminal repeat (ITR). In some embodiments, the at least one ITR comprises a 5’ ITR and a 3’ ITR. In some embodiments, the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto. In some embodiments, the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17. In some embodiments, the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical thereto. In some embodiments, the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21.Attorney Docket No. 14640.0304-00304

[0037] In some embodiments, the recombinant viral genome further comprises a nucleotide sequence encoding one or more microRNA (miR) binding sites, optionally wherein the one or more miR binding sites reduces or prevents expression of CDKL5 in dorsal root ganglia. In some embodiments, the one or more miR binding sites comprises one, two, three, or four miR183 binding sites. In some embodiments, the recombinant viral genome comprises a nucleotide sequence encoding four miR183 binding sites, optionally wherein the four miR183 binding sites are identical. In some embodiments, each of the four miR183 binding sites is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 6. In some embodiments, each of the four miR183 binding sites is encoded by the nucleotide sequence of SEQ ID NO: 6. In some embodiments, the miR183 binding sites are separated by a spacer, optionally wherein the spacer is encoded by the nucleotide sequence GATAGTTA.

[0038] In some embodiments, the recombinant viral genome further comprises a nucleotide sequence encoding a microRNA183 (miR183) binding site series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 7 or a nucleotide 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 thereto. In some embodiments, the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the nucleotide sequence encoding the miR183 binding site series consists of the nucleotide sequence of SEQ ID NO: 7.

[0039] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) a 5’ inverted terminal repeat (ITR); (b) a promoter; (c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence; (d) a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE); (e) a polyadenylation (poly A) sequence; and (f) a 3’ ITR.

[0040] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto; (c) the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide 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 thereto; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto; (e) the polyA sequence comprises the nucleotide sequence of SEQ IDAttorney Docket No. 14640.0304-00304NO: 20 or a nucleotide 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 thereto; and / or (f) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

[0041] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto; (c) the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide 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 thereto; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

[0042] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto; (c) the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.Attorney Docket No. 14640.0304-00304

[0043] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto; (c) the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

[0044] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18; (c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

[0045] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18; (c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.Attorney Docket No. 14640.0304-00304

[0046] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18; (c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21.

[0047] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18; (c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21.

[0048] In some embodiments, the recombinant viral genome comprises, in 5’ to 3’ order: (a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17; (b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18; (c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16; (d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19; (e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20; and (f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21.

[0049] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22 or a nucleotide 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 thereto. In some embodiments, the recombinant viral genome comprises a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 22.

[0050] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22.

[0051] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23.

[0052] In some embodiments, the recombinant viral genome further comprises a nucleotide sequence encoding one or more microRNA (miR) binding sites, optionally wherein the one orAttorney Docket No. 14640.0304-00304more miR binding sites reduces or prevents expression of CDKL5 in dorsal root ganglia. In some embodiments, the nucleotide sequence encoding the one or more miR binding sites comprises the nucleotide sequence of SEQ ID NO: 6. In some embodiments, the nucleotide sequence encoding the one or more miR binding sites encodes four miR binding sites, wherein each of the four miR binding sites is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 6. In some embodiments, the recombinant viral genome further comprises a nucleotide sequence encoding a microRNA (miR) binding site series, wherein the nucleotide sequence encoding the miR binding site series comprises the nucleotide sequence of SEQ ID NO: 7.

[0053] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

[0054] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising a recombinant viral genome and an AAV9 capsid variant, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 1; (ii) the amino acid sequence of SEQ ID NO: 24; and / or (iii) the amino acid sequence of SEQ ID NO: 25; and wherein the recombinant viral genome comprises: (a) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 17; (b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18; (c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19; (e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site series comprises the nucleotide sequence of SEQ ID NO: 6; (f) a polyadenylation (poly A) sequence comprising the nucleotide sequence of SEQ ID NO: 20; and (g) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

[0055] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

[0056] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising a recombinant viral genome and an AAV9 capsid variant, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 4; (ii) the amino acid sequence of SEQ ID NO: 26; and / or (iii) the amino acid sequence of SEQ ID NO: 27; and wherein the recombinant viral genome comprises: (a) a 5’ inverted terminal repeat (5’Attorney Docket No. 14640.0304-00304ITR) comprising the nucleotide sequence of SEQ ID NO: 17; (b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18; (c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19; (e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6; (f) a polyadenylation (poly A) sequence comprising the nucleotide sequence of SEQ ID NO: 20; and (g) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

[0057] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

[0058] In some embodiments, the present disclosure provides an adeno-associated virus (AAV) particle comprising a recombinant viral genome and an AAV9 capsid variant, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 45; (ii) the amino acid sequence of SEQ ID NO: 46; and / or (iii) the amino acid sequence of SEQ ID NO: 47; and wherein the recombinant viral genome comprises: (a) a 5’ inverted terminal repeat (5’ ITR) comprising the nucleotide sequence of SEQ ID NO: 17; (b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18; (c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19; (e) optionally a nucleotide sequence encoding a microRNA binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6; (f) a polyadenylation (poly A) sequence comprising the nucleotide sequence of SEQ ID NO: 20; and (g) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

[0059] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

[0060] In some embodiments, the present disclosure provides a cell comprising the AAV particle described 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.Attorney Docket No. 14640.0304-00304

[0061] In some embodiments, the present disclosure provides a method of making the AAV particle described herein, the method comprising: (i) providing a cell comprising the recombinant viral genome comprising a CDKL5-encoding sequence and a nucleic acid encoding the AAV9 capsid variant; and (ii) incubating the cell under conditions suitable to encapsulate the recombinant viral genome in the AAV9 capsid variant; thereby making the AAV particle.

[0062] In some embodiments, the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 1; (ii) the amino acid sequence of SEQ ID NO: 24; and / or (iii) the amino acid sequence of SEQ ID NO: 25; and the recombinant viral genome comprises: (a) a 5’ITR comprising the nucleotide sequence of SEQ ID NO: 17; (b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18; (c) a CDKL5 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) a WPRE comprising the nucleotide sequence of SEQ ID NO: 19; (e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6; (f) a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and (g) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

[0063] In some embodiments, the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 4; (ii) the amino acid sequence of SEQ ID NO: 26; and / or (iii) the amino acid sequence of SEQ ID NO: 27; and the recombinant viral genome comprises: (a) a 5’ITR comprising the nucleotide sequence of SEQ ID NO: 17; (b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18; (c) a CDKL5 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; (d) a WPRE comprising the nucleotide sequence of SEQ ID NO: 19; (e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6; (f) a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and (g) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

[0064] In some embodiments, the present disclosure provides the AAV9 capsid variant comprises: (i) the amino acid sequence of SEQ ID NO: 45; (ii) the amino acid sequence of SEQ ID NO: 46; and / or (iii) the amino acid sequence of SEQ ID NO: 47; and the recombinant viral genome comprises: (a) a 5’ITR comprising the nucleotide sequence of SEQ ID NO: 17; (b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18; (c) a CDKL5 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQAttorney Docket No. 14640.0304-00304ID NO: 16; (d) a WPRE comprising the nucleotide sequence of SEQ ID NO: 19; (e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6; (f) a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and (g) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

[0065] In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

[0066] In some embodiments, the method of making further comprises, prior to step (i), introducing a nucleic acid comprising the recombinant viral genome into the cell. In some embodiments, the method of making further comprises, prior to step (i), introducing the nucleic acid encoding the AAV9 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 Sf9 cell), or a bacterial cell.

[0067] In some embodiments, the present disclosure provides a pharmaceutical composition comprising the AAV particle described herein and a pharmaceutically acceptable excipient.

[0068] In some embodiments, the present disclosure provides a method of delivering an AAV particle encoding cyclin-dependent kinase-like 5 (CDKL5) to a cell, comprising administering an effective amount of the AAV particle described herein or the pharmaceutical composition described herein. In some embodiments, the cell is in a subject. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having a CDKL5-related disorder.

[0069] In some embodiments, the present disclosure provides a method of treating a subject having or diagnosed with having a CDKL5-related disorder, or at least one symptom thereof, comprising administering to the subject an effective amount of the AAV particle described herein or the pharmaceutical composition described herein. In some embodiments, the CDKL5-related disorder is a CDKL5-related neurodegenerative or neuromuscular disorder. In some embodiments, the CDKL5-related neurodegenerative or neuromuscular disorder is CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome. In some embodiments, the present disclosure provides a method of treating a subject having or having or diagnosed with having a CDKL5-related disorder, or treating at least one symptom thereof, wherein the CDKL5-related disorder is CDKL5Attorney Docket No. 14640.0304-00304deficiency disorder (CDD), comprising administering to the subject an effective amount of the AAV particle described herein or the pharmaceutical composition described herein.

[0070] In some embodiments, the subject has one or more mutations in a CDKL5 gene. In some embodiments, the subject has a reduced level of CDKL5 activity as compared to a reference level in an individual who does not have a CDKL5-related disorder. In some embodiments, the treating results in prevention or progression of the disorder or at least one symptom thereof in the subject. In some embodiments, the treating results in amelioration of at least one symptom of the disorder in the subject, e.g., as indicated by one or more biomarkers. In some embodiments, the one or more biomarkers comprises neurofilament light chain or a marker of CDKL5 activity, e.g., as measured by phosphorylation levels of substrate proteins, e.g., MECP2, or as measured by mass spectrometry.

[0071] In some embodiments, the at least one symptom comprises epilepsy (e.g., early-onset epilepsy), autism, deficits in cognition, limited motor skills, sleep difficulties, visual impairment, low muscle tone, gastrointestinal reflux, behavioral symptoms (including episodes of laughing or crying that occur for what appears to be no reason, hypersensitivity to touch, and disrupted sleep), facial appearance changes (including microcephaly; a high, broad forehead; large, deep-set eyes; smaller-than-normal space between the nose and upper lip; an upturned nose; full lips and / or widely-spaced teeth), difficulties standing and walking, small, cold feet, lack of or poor eye contact, frequent sideways glances, cortical visual impairment or cortical blindness, bruxism, limited or absent speech, difficulties eating, stereotypies, limited ability to make small and / or focused hand movements, gastroesophageal reflux, constipation, or a combination thereof.

[0072] In some embodiments, the subject is a human.

[0073] In some embodiments, the AAV particle or the pharmaceutical composition is delivered to a cell, tissue, or region of the central nervous system (CNS) of the subject. In some embodiments, the cell, tissue, or region of the CNS comprises: the parenchyma, cortex, substantia nigra, caudate cerebellum, striatum, corpus callosum, cerebellum, brain stem, caudate-putamen, thalamus, hippocampus, superior colliculus, spinal cord, or a combination thereof and / or neurons (e.g. glutamatergic neurons, GABAergic neurons, or a combination thereof). In some embodiments, the cell, tissue, or region of the CNS is a cell, tissue, or region of the cortex (e.g., motor cortex), hippocampus, striatum, thalamus, or cerebellum, and / or wherein the cell of the CNS is a glutamatergic neuron and / or GABAergic neuron.

[0074] In some embodiments, the AAV particle or pharmaceutical composition is delivered to the subject via intravenous administration.Attorney Docket No. 14640.0304-00304

[0075] In some embodiments, the method of delivering or treating further comprises evaluating, e.g., measuring, the level of CDKL5 gene expression, CDKL5 mRNA expression, and / or CDKL5 protein expression in the subject, e.g., in a cell, tissue, or fluid of the subject. In some embodiments, the level of CDKL5 protein expression is measured by an enzyme-linked immunosorbent assay (ELISA), a Western blot, or an immunohistochemistry assay. In some embodiments, evaluating the level of CDKL5 gene, mRNA, and / or protein expression is performed before and after administering the AAV particle or pharmaceutical composition, optionally wherein the subject’s level of CDKL5 gene, mRNA, and / or protein expression before administration is compared to the subject’s level of CDKL5 gene, mRNA, and / or protein expression after administration.

[0076] In some embodiments, the present disclosure provides the level of CDKL5 gene, mRNA, and / or protein expression is evaluated in a cell or tissue of the CNS in the subject. In some embodiments, the cell or tissue of the CNS comprises: the parenchyma, cortex, substantia nigra, caudate cerebellum, striatum, corpus callosum, cerebellum, brain stem, caudate-putamen, thalamus, hippocampus, superior colliculus, spinal cord, or a combination thereof and / or neurons (e.g. glutamatergic neurons, GABAergic neurons, or a combination thereof). In some embodiments, the cell or tissue of the CNS is a cell or tissue of the cortex (e.g., motor cortex), hippocampus, striatum, thalamus, or cerebellum, and / or wherein the cell of the CNS is a glutamatergic neuron and / or GABAergic neuron.

[0077] In some embodiments, the subject’s level of CDKL5 protein expression after administration of the AAV particle or pharmaceutical composition is increased relative to the subject’s level of CDKL5 protein expression before administration of the AAV particle or pharmaceutical composition.

[0078] In some embodiments, the method of treating or delivering further comprises evaluating, e.g., measuring, the level of CDKL5 protein activity in the subject, e.g., in a cell or tissue of the subject. In some embodiments, the method of treating or delivering further comprises administering the AAV particle or pharmaceutical composition to the subject results in an increase in: (i) the level of CDKL5 activity in a cell, tissue, or fluid (e.g., a cell or tissue of the CNS, e.g., the cortex, hippocampus, striatum, thalamus, cerebellum, glutamatergic neurons, GABAergic neurons, or a combination thereof) of the subject, relative to baseline and / or relative to the level of CDKL5 activity in a cell, tissue, or fluid of an individual with a CDKL5-related disorder who has not been administered the AAV particle or pharmaceutical composition; (ii) the number and / or level of recombinant viral genomes (VG) per cell level in a cell or tissue of the CNS (e.g., the cortex, hippocampus, striatum, thalamus,Attorney Docket No. 14640.0304-00304cerebellum, glutamatergic neurons, GABAergic neurons, or a combination thereof) of the subject, relative to the number and / or level of VG per cell in a peripheral cell or tissue of the subject; and / or (iii) the level of CDKL5 protein expression, CDKL5 mRNA expression, or CDKL5 gene expression in a cell or tissue (e.g., a cell or tissue of the CNS, e.g., the cortex, hippocampus, striatum, thalamus, cerebellum, glutamatergic neurons, GABAergic neurons, or a combination thereof) of the subject relative to baseline and / or relative to the level of CDKL5 protein expression, CDKL5 mRNA expression, or CDKL5 gene expression in a cell or tissue of an individual with a CDKL5-related disorder who has not been administered the AAV particle or pharmaceutical composition.

[0079] In some embodiments, the method of delivering or treating further comprises administering to the subject at least one additional agent and / or therapy suitable for treating the CDKL5-related disorder or at least one symptom thereof. In some embodiments, the at least one additional agent and / or therapy comprises one or more anti-epileptic drugs (e.g., bromide, clobazam, felbamate, ganaxolone, lamotrigine, levetiracetam, phenobarbital, topiramate, valproate, or a combination thereof).

[0080] In some embodiments, the method of delivering or treating further comprises administering an immunosuppressant to the subject. In some embodiments, the immunosuppressant comprises adrenocorticotropic hormone, a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), eculizumab hydroxychloroquine, mycophenolate mofetil, rapamycin, rituximab, and / or tacrolimus.

[0081] In some embodiments, the present disclosure provides the AAV particle described herein or the pharmaceutical composition described herein, for use in the treatment of a CDKL5-related disorder or at least one symptom thereof in a subject; optionally wherein the CDKL5-related disorder is CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome.

[0082] In some embodiments, the present disclosure provides the AAV particle or pharmaceutical composition described herein, wherein the subject has, has been diagnosed with having, or is at risk of having the CDKL5-related disorder or at least one symptom thereof; optionally wherein the CDKL5-related disorder is CDD, developmental and epileptic encephalopathy 2, or atypical Rett syndrome.

[0083] In some embodiments, the present disclosure provides use of the AAV particle described herein, or the pharmaceutical composition described herein, in the manufacture of a medicament for the treatment of an CDKL5-related disorder or at least one symptom thereof in a subject; optionally wherein the CDKL5-related disorder is CDKL5 deficiency disorderAttorney Docket No. 14640.0304-00304(CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome. In some embodiments, the subject has, has been diagnosed with having, or is at risk of having the CDKL5-related disorder or at least one symptom thereof; optionally wherein the CDKL5-related disorder is CDD, developmental and epileptic encephalopathy 2, or atypical Rett syndrome.

[0084] In some embodiments, the present disclosure provides the AAV particle described herein or the pharmaceutical composition described herein for use in a method of treating a disorder described herein.Brief Description of the Drawings

[0085] FIGs. 1A-1E depict vector genome (VG) quantification (VG / diploid genome (DNA)) for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 8 (SEQ ID NO: 38), Construct 1 (SEQ ID NO: 22), Construct 4 (SEQ ID NO: 39), Construct 7 (SEQ ID NO: 29), or Construct 6 (SEQ ID NO: 41). FIG. 1A depicts expression levels of human CDKL5 (hCDKL5) DNA relative to P-actin. FIG. IB depicts expression levels ofWPRE DNA relative to P-actin. FIG. 1C depicts expression levels of hCDKL5 DNA relative to mouse Cdkl5 (mCdkl5). FIG. ID depicts expression levels ofWPRE DNA relative to mCdkl5. FIG. IE depicts expression levels of mCdkl5 DNA relative to P-actin.

[0086] FIGs. 2A-2E depict transgene expression levels measured by RT-qPCR analysis of bulk RNA for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 8 (SEQ ID NO: 38), Construct 1 (SEQ ID NO: 22), Construct 4 (SEQ ID NO: 39), Construct 7 (SEQ ID NO: 29), or Construct 6 (SEQ ID NO: 41). FIG. 2A depicts the mRNA levels of hCDKL5 relative to P-actin. FIG. 2B depicts the mRNA levels ofWPRE relative to P-actin. FIG. 2C depicts the mRNA levels of hCDKL5 relative to mCdkl5. FIG. 2D depicts the mRNA levels ofWPRE relative to mCdkl5. FIG. 2E depicts the mRNA levels of mCdkl5 relative to P-actin.

[0087] FIGs. 3A-3D depict the percentage of neurons that were hCDKL5-positive for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 5 (SEQ ID NO: 42), or Construct 8 (SEQ ID NO: 38), where a cell was considered hCDKL5 -positive if the hCDKL5 fluorescent area was greater than 0 pm2. FIG.3A depicts the percentage of hCDKL5-positive neurons in the cortex. FIG. 3B depicts the percentage of hCDKL5-positive neurons in the CAI hippocampal region. FIG. 3C depicts the percentage of hCDKL5-positive neurons in the CA2 hippocampal region. FIG. 3D depicts the percentage of hCDKL5-positive neurons in the CA3 hippocampal region.Attorney Docket No. 14640.0304-00304

[0088] FIGs. 4A-4D depict the percentage of neurons that were hCDKL5-positive for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 5 (SEQ ID NO: 42), or Construct 8 (SEQ ID NO: 38), where a cell was considered hCDKL5-positive if the cell’s hCDKL5 fluorescent area was greater than the 25thpercentile of mCdkl5 fluorescent area. FIG. 4A depicts the percentage of hCDKL5-positive neurons in the cortex. FIG. 4B depicts the percentage of hCDKL5 -positive neurons in the CAI hippocampal region. FIG. 4C depicts the percentage of hCDKL5 -positive neurons in the CA2 hippocampal region. FIG. 4D depicts the percentage of hCDKL5-positive neurons in the CA3 hippocampal region.

[0089] FIGs. 5A-5D show the percentage of neurons that were mCdkl 5 -positive for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 5 (SEQ ID NO: 42), or Construct 8 (SEQ ID NO: 38), where a cell was considered mCdkl5-positive if the cell’s mCdkl5 fluorescent area was greater than 0 pm2. FIG. 5A depicts the percentage of mCdkl5-positive neurons in the cortex. FIG. 5B depicts the percentage of mCdkl5-positive neurons in the CAI hippocampal region. FIG. 5C depicts the percentage of mCdkl5-positive neurons in the CA2 hippocampal region. FIG. 5D depicts the percentage of mCdkl5-positive neurons in the CA3 hippocampal region.

[0090] FIGs. 6A-6D depict the percentage of mCdkl5-positive neurons that were hCDKL5-positive (i.e., the percentage of mCdkl5 / NeuN double-stained neurons that were hCDKL5 / mCdkl5 / NeuN triple stained) for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 5 (SEQ ID NO: 42), or Construct 8 (SEQ ID NO: 38). FIG. 6A depicts the percentage of mCdkl5-positive neurons that were hCDKL5-positive in the cortex. FIG. 6B depicts the percentage of mCdkl5-positive neurons that were hCDKL5 -positive in the CAI hippocampal region. FIG.6C depicts the percentage of mCdkl 5 -positive neurons that were hCDKL5 -positive in the CA2 hippocampal region. FIG. 6D depicts the percentage of mCdkl5-positive neurons that were hCDKL5 -positive in the CA3 hippocampal region.

[0091] FIGs. 7A-7D depict the percentage of neurons expressing WPRE for mice at 4-weeks post-IV injection of PBS or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22) or Construct 7 (SEQ ID NO: 29). FIG. 7A depicts the percentage of neurons with WPRE mRNA in the cortex. FIG. 7B depicts the percentage of neurons with WPRE mRNA in the CAI hippocampal region. FIG. 7C depicts the percentage of neurons with WPRE mRNA in the CA2 hippocampal region. FIG. 7D depicts the percentage of neurons with WPRE mRNA in the CA3 hippocampal region.Attorney Docket No. 14640.0304-00304

[0092] FIGs. 8A-8E depict CDKL5 transgene expression in the hippocampus (HPC), cortex (CTX), striatum (STR), and thalamus (Thai) 4 weeks after dosing mice via IV injection with PBS (Veh) or PHP.eB viral particles (denoted as AAV) carrying Construct 1 (SEQ ID NO: 22). FIG. 8A depicts mCdkl5 expression in a wildtype (WT) mouse. FIG. 8B depicts hCDKL5 expression in a mouse administered PHP.eB-Construct 1. FIG. 8C depicts a lack of hCDKL5 expression in a PBS-treated mouse. The percentage of neurons positive for mCDKL5 or hCDKL5 for mice treated with PBS or PHP.eB-Construct 1 are reported in FIG.8D (mCDKL5) and FIG. 8E (hCDKL5). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0093] FIGs. 9A-9B depict transgene expression driven by the human synapsin (hSynl) promoter in Construct 1 (SEQ ID NO: 22). FIG. 9 A shows the injection site in the thalamus and FIG. 9B depicts hCDKL5 expression at the injection compared to a contralateral site 19 days post-injection.

[0094] FIGs. 10A-10C depict neuronal expression of hCDKL5 following injection with PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 10A depicts hCDKL5 staining. FIG. 10B depicts NeuN staining. FIG. 10C depicts colocalization of hCDKL5 and NeuN staining.

[0095] FIG. HA depicts DAPI staining of neurons at an injection site following administration of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 11B depicts hCDKL5 -positive cells in the injection site. FIG. 11C depicts DAPI staining of a contralateral site. FIG. 11D depicts a lack of hCDKL5-positive cells in the contralateral site.

[0096] FIGs. 12A-12E depict expression results in the cerebellum 4 weeks following IV injection of wildtype or CDKL5-KO mice with PBS or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 12A depicts vector genome copy. FIG. 12B depicts hCDKL5 mRNA. FIG. 12C depicts the percentage of hCDKL5-positive neurons in the cortex and hippocampus, as measured by in situ hybridization. FIG. 12D depicts CDKL5 protein concentration. FIG. 12E depicts kinase activity of the CDKL5 protein. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0097] FIGs. 13A and 13B depict the effects of administration of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 4 (SEQ ID NO: 39), or Construct 8 (SEQ ID NO: 38) in wildtype and CDKL5-KO mice. FIG. 13A depicts the occurrence of sudden unexpected death in epilepsy (SUDEP). FIG. 13B depicts the severity of seizures as measured using the Racine scale. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001

[0098] FIGs. 14A-14F depict effects on motor function in CDKL5-KO mice when a WPRE or Myc tag is included in payloads. FIG. 14A depicts aggregate scores of hindlimb claspingAttorney Docket No. 14640.0304-00304behavior. FIG. 14B depicts individual scores for wildtype (WT) mice administered PBS (Veh). FIG. 14C depicts individual scores for CDKL5-K0 mice administered PBS (Veh). FIG. 14D depicts individual scores for CDKL5-K0 mice administered PHP.eB viral particles carrying Construct 4 (SEQ ID NO: 39). FIG. 14E depicts individual scores for CDKL5-K0 mice administered PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 14F depicts individual scores for CDKL-KO mice administered PHP.eB viral particles carrying Construct 8 (SEQ ID NO: 38). Wk = week. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0099] FIGs. 15A-15B depict results from recordings of ex vivo neural hyperactivity in mice administered PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 15A depicts a whole cell patch clamp in the CA2 region of the hippocampus from a mouse. FIG.15B depicts a comparison of the decay time of NMDA current in CDKL5-KO mice as compared to wildtype (WT) mice following administration of PBS (Veh) or PHP.eB viral particles carrying Construct 1 (AAV). *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0100] FIGs. 16A-16B depict a reduction in seizure susceptibility and mortality in CDKL5 KO mice following administration of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 4 (SEQ ID NO: 39), or Construct 8 (SEQ ID NO: 38). FIG. 16A depicts percent mortality. FIG. 16B depicts maximum seizure scores. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0101] FIGs. 17A-17B depict expression of CDKL5 in CDKL5-KO mice dosed with vehicle or PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22), Construct 4 (SEQ ID NO: 39), or Construct 8 (SEQ ID NO: 38). FIG. 17A depicts the hCDKL5 in total protein (pg / g). FIG. 17B depicts hCDKL5 in total protein, measured as a percentage of hCDKL5 protein in the wildtype (WT) mice dosed with vehicle. *P<0.05, **P<0.01, ***P<0.001, ****p<0.0001.

[0102] FIGs. 18A-18E depict effects of various doses of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22) on CDKL5 KO mice. FIG. 18A depicts mortality during seizures. FIG. 18B depicts the latency to death. FIG. 18C depicts the hindlimb clasping behavior score before dosing. FIG. 18D depicts the hindlimb clasping behavior score after dosing. FIG. 18E depicts a summary of hindlimb clasping behavior scores. Veh = Vehicle; WT = wildtype. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0103] FIG. 19A depicts vector genome copies per diploid genome in CDKL5-KO mouse cerebrum following administration of various doses of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 19B depicts normalized CDKL5 protein expression in CDKL5-KO mouse cerebrum following administration of various doses of PHP.eB viralAttorney Docket No. 14640.0304-00304particles carrying Construct 1 (SEQ ID NO: 22). FIG. 19C depicts the percentage of neurons in CDKL5-K0 mouse cortex that are hCDKL5-positive following administration of various doses of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 19D depicts the percentage of neurons in CDKL5-K0 mouse hippocampus that are hCDKL5-positive following administration of various doses of PHP.eB viral particles carrying Construct 1 (SEQ ID NO: 22). Veh = Vehicle; WT = wildtype.

[0104] FIGs. 20A-20D depict results at 4-weeks post-IV injection of PBS or TTM-027 viral particles carrying Construct 1 (SEQ ID NO: 22) in wildtype (WT) or CDKL5-KO mice. FIG.20A depicts the percentage of neurons that are hCDKL5-positive in the cortex. FIG. 20B depicts the percentage of neurons that are hCDKL5-positive in the hippocampus. FIG. 20C depicts hindlimb clasping behavior for all groups of mice after dosing. FIG. 20D depicts hindlimb clasping behavior before and after dosing. *P<0.05, **P<0.01, ***P<0.001, ****p<0.0001.

[0105] FIGs. 21A-21D depict survival and latency to seizure data obtained from either wildtype (WT) or CDKL5-KO mice at 4-weeks post-IV injection of PBS or TTM-027 viral particles carrying Construct 1 (SEQ ID NO: 22). FIG. 21A depicts probability of survival (KM survival analysis). FIG. 21B depicts percent mortality during seizure. FIG. 21C depicts latency to death. FIG. 21D depicts the latency to seizure.

[0106] FIG. 22 depicts CDKL5 protein expression in wildtype C57 mice at 4-weeks post-IV injection of vehicle-PBS, PHP.eB, or TTM-027 viral particles carrying Construct 1 (SEQ ID NO: 22) or Construct 8 (SEQ ID NO: 38).Detailed DescriptionI. CompositionsA. Nucleic Acids and Viral Genomes1. CDKL5-Encoding Sequence

[0107] In some embodiments, the present disclosure provides a nucleic acid (e.g., an isolated nucleic acid) comprising a polynucleotide encoding CDKL5, also referred to as a CDKL5-encoding sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) is or is comprised within a viral genome (e.g., recombinant viral genome). Accordingly, in some embodiments, the present disclosure provides an isolated nucleic acid comprising a CDKL5-encoding sequence and, in some embodiments, the present disclosure provides a viral genome (e.g., recombinant viral genome) comprising said isolated nucleic acid.Attorney Docket No. 14640.0304-00304

[0108] In some embodiments, the CDKL5 -encoding sequence encodes human CDKL5. In some embodiments, the CDKL5 -encoding sequence encodes wildtype CDKL5. In some embodiments, the CDKL5 -encoding sequence encodes a CDKL5 isoform.

[0109] In some embodiments, the CDKL5 -encoding sequence encodes CDKL5 isoform 1 (e.g., the CDKL5 amino acid sequence of SEQ ID NO: 34), CDKL5 isoform 2 (e.g., the CDKL5 amino acid sequence of SEQ ID NO: 35), CDKL5 isoform CRA A (e.g., the CDKL5 amino acid sequence of SEQ ID NO: 36), or CDKL5 isoform CRA b (e.g., the CDKL5 amino acid sequence of SEQ ID NO: 37). In some embodiments, the CDKL5 comprises the amino acid sequence of any one of SEQ ID NOs: 34-37 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 thereto. In some embodiments, the CDKL5 comprises the amino acid sequence of any one of SEQ ID NOs: 34-37.

[0110] In some embodiments, the CDKL5 -encoding sequence encodes a CDKL5 protein comprising the amino acid sequence of SEQ ID NO: 14 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 thereto. In some embodiments, the CDKL5 protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the CDKL5 protein comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the CDKL5 protein comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the CDKL5 protein comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the CDKL5 protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the CDKL5 protein comprises the amino acid sequence of SEQ ID NO: 14. In some embodiments, the CDKL5 protein consists of the amino acid sequence of SEQ ID NO: 14.

[0111] In some embodiments, the CDKL5 -encoding sequence comprises a human CDKL5 nucleotide sequence. In some embodiments, the CDKL5 -encoding sequence comprises a wildtype CDKL5 nucleotide sequence. In some embodiments, the CDKL5-encoding sequence comprises a codon-optimized CDKL5 nucleotide sequence. In some embodiments, the CDKL5-encoding sequence comprises one or more CpG motifs. In some embodiments,Attorney Docket No. 14640.0304-00304the CDKL5 -encoding sequence is CpG-depleted. In some embodiments, the CDKL5-encoding sequence comprises no CpG motifs (i.e., is CpG-free).

[0112] In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide 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 thereto. In some embodiments, the CDKL5-encoding sequence comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 15. In some embodiments, the CDKL5 -encoding sequence comprises a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO: 15. In some embodiments, the CDKL5-encoding sequence comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of SEQ ID NO: 15. In some embodiments, the CDKL5 -encoding sequence comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of SEQ ID NO: 15. In some embodiments, the CDKL5 -encoding sequence comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 15. In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15. In some embodiments, the CDKL5 -encoding sequence consists of the nucleotide sequence of SEQ ID NO: 15.

[0113] In some embodiments, the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16 or a nucleotide 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 thereto. In some embodiments, the CDKL5-encoding sequence comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the CDKL5 -encoding sequence comprises a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the CDKL5-encoding sequence comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the CDKL5 -encoding sequence comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the CDKL5 -encoding sequence comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16. In some embodiments, the CDKL5 -encoding sequence consists of the nucleotide sequence of SEQ ID NO: 16.Attorney Docket No. 14640.0304-00304

[0114] Non-limiting examples of CDKL5 amino acid sequences are provided in Table 1.

[0115] Non-limiting examples of CDKL5 -encoding sequences are provided in Table 2A.

[0116] The present disclosure provides nucleic acids (e.g., an isolated nucleic acids) or viral genomes (e.g., recombinant viral genomes) encoding a Myc-tagged CDKL5 protein. In some embodiments, the tag is a Myc tag. In some embodiments, the Myc tag is linked to the CDKL5 protein by a GGGS (SEQ ID NO: 44) spacer sequence. In some embodiments, the Myc tag and spacer (e.g., of amino acid sequence EQKLISEEDLGGGGS (SEQ ID NO: 31)) is located immediately after the N-terminal methionine of the encoded CDKL5. Table 2B provides exemplary nucleotide sequences that encode Myc-tagged CDKL5.Table 1. Exemplary CDKL5 Amino Acid SequencesSEQ Amino Acid SequenceID NO.14 MKIPNIGNVMNKFEILGWGEGAYGWLKCRHKETHEIVAIKKFKDSEENEEVKETTLRELKMLRTLKQE NIVELKEAFRRRGKLYLVFEYVEKNMLELLEEMPNGVPPEKVKSYIYQLIKAIHWCHKNDIVHRDIKPEN LLISHNDVLKLCDFGFARNLSEGNNANYTEYVATRWYRSPELLLGAPYGKSVDMWSVGCILGELSDGQPL FPGESEIDQLFTIQKVLGPLPSEQMKLFYSNPRFHGLRFPAVNHPQSLERRYLGILNSVLLDLMKNLLKL DPADRYLTEQCLNHPTFQTQRLLDRSPSRSAKRKPYHVESSTLSNRNQAGKSTALQSHHRSNSKDIQNLS VGLPRADEGLPANESFLNGNLAGASLSPLHTKTYQASSQPGSTSKDLTNNNIPHLLSPKEAKSKTEFDFN IDPKPSEGPGTKYLKSNSRSQQNRHSFMESSQSKAGTLQPNEKQSRHSYIDTIPQSSRSPSYRTKAKSHG ALSDSKSVSNLSEARAQIAEPSTSRYFPSSCLDLNSPTSPTPTRHSDTRTLLSPSGRNNRNEGTLDSRRT TTRHSKTMEELKLPEHMDSSHSHSLSAPHESFSYGLGYTSPFSSQQRPHRHSMYVTRDKVRAKGLDGSLS IGQGMAARANSLQLLSPQPGEQLPPEMTVARSSVKETSREGTSSFHTRQKSEGGVYHDPHSDDGTAPKEN RHLYNDPVPRRVGSFYRVPSPRPDNSFHENNVSTRVSSLPSESSSGTNHSKRQPAFDPWKSPENISHSEQ LKEKEKQGFFRSMKKKKKKSQTVPNSDSPDLLTLQKSIHSASTPSSRPKEWRPEKISDLQTQSQPLKSLR KLLHLSSASNHPASSDPRFQPLTAQQTKNSFSEIRIHPLSQASGGSSNIRQEPAPKGRPALQLPGQMDPG WHVSSVTRSATEGPSYSEQLGAKSGPNGHPYNRTNRSRMPNLNDLKETAL34 MKIPNIGNVMNKFEILGWGEGAYGWLKCRHKETHEIVAIKKFKDSEENEEVKETTLRELKMLRTLKQE NIVELKEAFRRRGKLYLVFEYVEKNMLELLEEMPNGVPPEKVKSYIYQLIKAIHWCHKNDIVHRDIKPEN LLISHNDVLKLCDFGFARNLSEGNNANYTEYVATRWYRSPELLLGAPYGKSVDMWSVGCILGELSDGQPL FPGESEIDQLFTIQKVLGPLPSEQMKLFYSNPRFHGLRFPAVNHPQSLERRYLGILNSVLLDLMKNLLKL DPADRYLTEQCLNHPTFQTQRLLDRSPSRSAKRKPYHVESSTLSNRNQAGKSTALQSHHRSNSKDIQNLS VGLPRADEGLPANESFLNGNLAGASLSPLHTKTYQASSQPGSTSKDLTNNNIPHLLSPKEAKSKTEFDFN IDPKPSEGPGTKYLKSNSRSQQNRHSFMESSQSKAGTLQPNEKQSRHSYIDTIPQSSRSPSYRTKAKSHG ALSDSKSVSNLSEARAQIAEPSTSRYFPSSCLDLNSPTSPTPTRHSDTRTLLSPSGRNNRNEGTLDSRRT TTRHSKTMEELKLPEHMDSSHSHSLSAPHESFSYGLGYTSPFSSQQRPHRHSMYVTRDKVRAKGLDGSLS IGQGMAARANSLQLLSPQPGEQLPPEMTVARSSVKETSREGTSSFHTRQKSEGGVYHDPHSDDGTAPKEN RHLYNDPVPRRVGSFYRVPSPRPDNSFHENNVSTRVSSLPSESSSGTNHSKRQPAFDPWKSPENISHSEQ LKEKEKQGFFRSMKKKKKKSQTVPNSDSPDLLTLQKSIHSASTPSSRPKEWRPEKISDLQTQSQPLKSLR KLLHLSSASNHPASSDPRFQPLTAQQTKNSFSEIRIHPLSQASGGSSNIRQEPAPKGRPALQLPDGGCDG RRQRHHSGPQDRRFMLRTTEQQGEYFCCGDPKKPHTPCVPNRALHRPISSPAPYPVLQVRGTSMCPTLQV RGTDAFSCPTQQSGFSFFVRHVMREALIHRAQVNQAALLTYHENAALTGK35 MKIPNIGNVMNKFEILGWGEGAYGWLKCRHKETHEIVAIKKFKDSEENEEVKETTLRELKMLRTLKQE NIVELKEAFRRRGKLYLVFEYVEKNMLELLEEMPNGVPPEKVKSYIYQLIKAIHWCHKNDIVHRDIKPEN LLISHNDVLKLCDFGFARNLSEGNNANYTEYVATRWYRSPELLLGAPYGKSVDMWSVGCILGELSDGQPL FPGESEIDQLFTIQKVLGPLPSEQMKLFYSNPRFHGLRFPAVNHPQSLERRYLGILNSVLLDLMKNLLKL DPADRYLTEQCLNHPTFQTQRLLDRSPSRSAKRKPYHVESSTLSNRNQAGKSTALQSHHRSNSKDIQNLS VGLPRADEGLPANESFLNGNLAGASLSPLHTKTYQASSQPGSTSKDLTNNNIPHLLSPKEAKSKTEFDFN IDPKPSEGPGTKYLKSNSRSQQNRHSFMESSQSKAGTLQPNEKQSRHSYIDTIPQSSRSPSYRTKAKSHG ALSDSKSVSNLSEARAQIAEPSTSRYFPSSCLDLNSPTSPTPTRHSDTRTLLSPSGRNNRNEGTLDSRRT TTRHSKTMEELKLPEHMDSSHSHSLSAPHESFSYGLGYTSPFSSQQRPHRHSMYVTRDKVRAKGLDGSLSIGQGMAARANSLQLLSPQPGEQLPPEMTVARSSVKETSREGTSSFHTRQKSEGGVYHDPHSDDGTAPKENAttorney Docket No. 14640.0304-00304SEQ Amino Acid SequenceID NO.RHLYNDPVPRRVGSFYRVPSPRPDNSFHENNVSTRVSSLPSESSSGTNHSKRQPAFDPWKSPENISHSEQ LKEKEKQGFFRSMKKKKKKSQTVPNSDSPDLLTLQKSIHSASTPSSRPKEWRPEKISDLQTQSQPLKSLR KLLHLSSASNHPASSDPRFQPLTAQQTKNSFSEIRIHPLSQASGGSSNIRQEPAPKGRPALQLPGQMDPG WHVSSVTRSATEGPSYSEQLGAKSGPNGHPYNRTNRSRMPNLNDLKETAL36 MELYLNADTRQETHEIVAIKKFKDSEENEEVKETTLRELKMLRTLKQENIVELKEAFRRRGKLYLVFEYV EKNMLELLEEMPNGVPPEKVKSYIYQLIKAIHWCHKNDIVHRDIKPENLLISHNDVLKLCDFGFARNLSE GNNANYTEYVATRWYRSPELLLGAPYGKSVDMWSVGCILGELSDGQPLFPGESEIDQLFTIQKVLGPLPS EQMKLFYSNPRFHGLRFPAVNHPQSLERRYLGILNSVLLDLMKNLLKLDPADRYLTEQCLNHPTFQTQRL LDRSPSRSAKRKPYHVESSTLSNRNQAGKSTALQSHHRSNSKDIQNLSVGLPRADEGLPANESFLNGNLA GASLSPLHTKTYQASSQPGSTSKDLTNNNIPHLLSPKEAKSKTEFDFNIDPKPSEGPGTKYLKSNSRSQQ NRHSFMESSQSKAGTLQPNEKQSRHSYIDTIPQSSRSPSYRTKAKSHGALSDSKSVSNLSEARAQIAEPS TSRYFPSSCLDLNSPTSPTPTRHSDTRTLLSPSGRNNRNEGTLDSRRTTTRHSKTMEELKLPEHMDSSHS HSLSAPHESFSYGLGYTSPFSSQQRPHRHSMYVTRDKVRAKGLDGSLSIGQGMAARANSLQLLSPQPGEQ LPPEMTVARSSVKETSREGTSSFHTRQKSEGGVYHDPHSDDGTAPKENRHLYNDPVPRRVGSFYRVPSPR PDNSFHENNVSTRVSSLPSESSSGTNHSKRQPAFDPWKSPENISHSEQLKEKEKQGFFRSMKKKKKKSQT VPNSDSPDLLTLQKSIHSASTPSSRPKEWRPEKISDLQTQSQPLKSLRKLLHLSSASNHPASSDPRFQPL TAQQTKNSFSEIRIHPLSQASGGSSNIRQEPAPKGRPALQLPDGGCDGRRQRHHSGPQDRRFMLRTTEQQ GEYFCCGDPKKPHTPCVPNRALHRPISSPAPYPVLQVRGTSMCPTLQVRGTDAFSCPTQQSGFSFFVRHV MREALIHRAQVNQAALLTYHENAALTGK37 MKIPNIGNVMNKFEILGWGEGAYGWLKCRHKETHEIVAIKKFKDSEENEEVKETTLRELKMLRTLKQE NIVELKEAFRRRGKLYLVFEYVEKNMLELLEEMPNGVPPEKVKSYIYQLIKAIHWCHKNDIVHRDIKPEN LLISHNDVLKLCDFGFARNLSEGNNANYTEYVATRWYRSPELLLGAPYGKSVDMWSVGCILGELSDGQPL FPGESEIDQLFTIQKVLGPLPSEQMKLFYSNPRFHGLRFPAVNHPQSLERRYLGILNSVLLDLMKNLLKL DPADRYLTEQCLNHPTFQTQRLLDRSPSRSAKRKPYHVESSTLSNRNQAGKSTALQSHHRSNSKDIQNLS VGLPRADEGLPANESFLNGNLAGASLSPLHTKTYQASSQPGSTSKDLTNNNIPHLLSPKEAKSKTEFDFN IDPKPSEGPGTKYLKSNSRSQQNRHSFMESSQSKAGTLQPNEKQSRHSYIDTIPQSSRSPSYRTKAKSHG ALSDSKSVSNLSEARAQIAEPSTSRYFPSSCLDLNSPTSPTPTRHSDTRTLLSPSGRNNRNEGTLDSRRT TTRHSKTMEELKLPEHMDSSHSHSLSAPHESFSYGLGYTSPFSSQQRPHRHSMYVTRDKVRAKGLDGSLS IGQGMAARANSLQLLSPQPGEQLPPEMTVARSSVKETSREGTSSFHTRQKSEGGVYHDPHSDDGTAPKEN RHLYNDPVPRRVGSFYRVPSPRPDNSFHENNVSTRVSSLPSESSSGTNHSKRQPAFDPWKSPENISHSEQ LKEKEKQGFFRSMKKKKKKSQTVPNSDSPDLLTLQKSIHSASTPSSRPKEWRPEKISDLQTQSQPLKSLR KLLHLSSASNHPASSDPRFQPLTAQQTKNSFSEIRIHPLSQASGGSSNIRQEPAPKGRPALQLPDGGCDG RRQRHHSGPQDRRFMLRTTEQQGEYFCCGDPKKPHTPCVPNRALHRPISSPAPYPVLQVRGTSMCPTLQVRGTDAFSCPTQQSGFSFFVRHVMREALIHRAQVNQAALLTYHENAALTGKTable 2A. Exemplary CDKL5-Encoding SequencesSEQ Nucleotide SequenceID NO.15 ATGAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGAGATCCTTGGGGTTGTAGGTGAAGGAGCC TATGGAGTTGTACTTAAATGCAGACACAAGGAAACACATGAAATTGTGGCGATCAAGAAATTCAAGGAC AGTGAAGAAAATGAAGAAGTCAAAGAAACGACTTTACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAG GAAAACATTGTGGAGTTGAAGGAAGCATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTT GAAAAAAATATGCTCGAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTAAAAAGCTAC ATCTATCAGCTAATCAAGGCTATTCACTGGTGCCATAAGAATGATATTGTCCATCGAGATATAAAACCA GAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGTGTGACTTTGGTTTTGCTCGTAATCTGTCA GAAGGCAATAATGCTAATTACACAGAGTACGTTGCCACCAGATGGTATCGGTCCCCAGAACTCTTACTT GGCGCTCCCTATGGAAAGTCCGTGGACATGTGGTCGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGA CAGCCTTTATTTCCTGGAGAAAGTGAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTT CCATCTGAGCAGATGAAGCTTTTCTACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCCAGCTGTTAAC CATCCTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTCTACTTGACCTAATGAAGAAT TTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAACAGTGTTTGAATCACCCTACATTTCAAACC CAGAGACTTCTGGATCGTTCTCCTTCAAGGTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAGCCGGCAAAAGTACTGCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACAttorney Docket No. 14640.0304-00304ATCCAGAACCTGAGTGTAGGCCTGCCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAAT GGAAACCTTGCTGGAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCAGCCAGCCTGGG TCTACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGCCCAAAAGAAGCCAAGTCAAAA ACAGAGTTTGATTTTAATATTGACCCAAAGCCTTCAGAAGGCCCAGGGACAAAGTACCTCAAGTCAAAC AGCAGATCTCAGCAGAACCGCCACTCATTCATGGAAAGCTCTCAAAGCAAAGCTGGGACACTGCAGCCC AATGAAAAGCAGAGTCGGCATAGCTATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGG ACCAAGGCCAAAAGCCATGGGGCACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCC CAAATTGCGGAGCCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAATTCTCCCACCAGC CCAACCCCCACCAGACACAGTGACACGAGAACTTTGCTCAGCCCTTCTGGAAGAAATAACCGAAATGAG GGAACGCTGGACTCACGTCGAACCACAACCAGACATTCTAAGACGATGGAGGAATTGAAGCTGCCGGAG CACATGGACAGTAGCCATTCCCATTCACTGTCTGCACCTCACGAATCTTTTTCTTATGGACTGGGCTAC ACCAGCCCCTTTTCTTCCCAGCAACGTCCTCATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGA GCCAAGGGCTTGGATGGAAGCTTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAACTC TTGTCACCCCAGCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTCGGTCAAAGAGACC TCCAGAGAAGGCACCTCTTCCTTCCATACACGCCAGAAGTCTGAGGGTGGAGTGTATCATGACCCACAC TCTGATGATGGCACAGCCCCCAAAGAAAATAGACACCTATACAATGATCCTGTGCCAAGGAGAGTTGGT AGCTTTTACAGAGTGCCATCTCCACGTCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTT TCTTCTCTACCATCAGAGAGCAGTTCTGGAACCAACCACTCAAAAAGACAACCAGCATTCGATCCATGG AAAAGTCCTGAAAATATTAGTCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAGGTCA ATGAAAAAGAAAAAGAAGAAATCTCAAACAGTACCCAATTCCGACAGCCCTGATCTTCTGACGTTGCAG AAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAGTGGCGCCCCGAGAAGATCTCAGAT CTGCAGACCCAAAGCCAGCCATTAAAATCACTGCGCAAGTTGTTACATCTCTCTTCGGCCTCAAATCAC CCGGCTTCCTCAGATCCCCGCTTCCAGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATT CGGATTCACCCCCTGAGCCAGGCCTCTGGCGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGC AGGCCAGCCCTCCAGCTGCCAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGAGTGCC ACAGAGGGCCCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGGCACCCCTATAACAGA ACAAATCGCTCACGAATGCCAAATCTGAATGATTTAAAAGAGACAGCCTTGTAA ATGAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGAGATCCTTGGGGTTGTAGGTGAAGGAGCC TATGGAGTTGTACTTAAATGCAGACACAAGGAAACACATGAAATTGTGGCCATCAAGAAATTCAAGGAC AGTGAAGAAAATGAAGAAGTCAAAGAAACCACTTTAAGGGAGCTTAAAATGCTTAGGACTCTCAAGCAG GAAAACATTGTGGAGTTGAAGGAAGCATTTAGGAGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTT GAAAAGAATATGCTAGAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTAAAAAGCTAC ATCTATCAGCTAATCAAGGCTATTCACTGGTGCCATAAGAATGATATTGTCCATAGGGATATAAAACCA GAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGTGTGACTTTGGTTTTGCTAGGAATCTGTCA GAAGGCAATAATGCTAATTACACAGAGTATGTTGCCACCAGATGGTATAGGTCCCCAGAACTCTTACTT GGAGCTCCCTATGGAAAGTCAGTGGACATGTGGTCTGTGGGCTGTATTCTTGGGGAGCTTAGTGATGGA CAGCCTTTATTTCCTGGAGAAAGTGAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTT CCATCTGAGCAGATGAAGCTTTTCTACAGTAATCCTAGGTTCCATGGGCTCAGGTTTCCAGCTGTTAAC CATCCTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTCTACTTGACCTAATGAAGAAT TTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAACAGTGTTTGAATCACCCTACATTTCAAACC CAGAGACTTCTGGATAGGTCTCCTTCAAGGTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACA TTGTCTAATAGAAACCAAGCAGGCAAAAGTACTGCTTTGCAGTCTCACCACAGATCTAACAGCAAGGAC ATCCAGAACCTGAGTGTAGGCCTGCCCAGGGCTGATGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAAT GGAAACCTTGCTGGAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCAGCCAGCCTGGG TCTACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGCCCAAAAGAAGCCAAGTCAAAA ACAGAGTTTGATTTTAATATTGACCCAAAGCCTTCAGAAGGCCCAGGGACAAAGTACCTCAAGTCAAAC AGCAGATCTCAGCAGAACAGGCACTCATTCATGGAAAGCTCTCAAAGCAAAGCTGGGACACTGCAGCCC AATGAAAAGCAGAGTAGGCATAGCTATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGG ACCAAGGCCAAAAGCCATGGGGCACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCC CAAATTGCTGAGCCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAATTCTCCCACCAGC CCAACCCCCACCAGACACAGTGACACCAGAACTTTGCTCAGCCCTTCTGGAAGAAATAACAGGAATGAG GGAACCCTGGACTCAAGGAGGACCACAACCAGACATTCTAAGACCATGGAGGAATTGAAGCTGCCTGAG CACATGGACAGTAGCCATTCCCATTCACTGTCTGCACCTCATGAATCTTTTTCTTATGGACTGGGCTAC ACCAGCCCCTTTTCTTCCCAGCAAAGGCCTCATAGGCATTCTATGTATGTGACCAGGGACAAAGTGAGA GCCAAGGGCTTGGATGGAAGCTTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAACTC TTGTCACCCCAGCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTCTGTCAAAGAGACC TCCAGAGAAGGCACCTCTTCCTTCCATACAAGGCAGAAGTCTGAGGGTGGAGTGTATCATGACCCACAC TCTGATGATGGCACAGCCCCCAAAGAAAATAGACACCTATACAATGATCCTGTGCCAAGGAGAGTTGGT AGCTTTTACAGAGTGCCATCTCCAAGGCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTT TCTTCTCTACCATCAGAGAGCAGTTCTGGAACCAACCACTCAAAAAGACAACCAGCATTTGATCCATGGAAAAGTCCTGAAAATATTAGTCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAGGTCAAttorney Docket No. 14640.0304-00304ATGAAAAAGAAAAAGAAGAAATCTCAAACAGTACCCAATTCAGACAGCCCTGATCTTCTGACCTTGCAG AAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAGTGGAGGCCAGAGAAGATCTCAGAT CTGCAGACCCAAAGCCAGCCATTAAAATCACTGAGGAAGTTGTTACATCTCTCTTCTGCCTCAAATCAC CCTGCTTCCTCAGATCCCAGGTTCCAGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATT AGGATTCACCCCCTGAGCCAGGCCTCTGGAGGGAGCAGCAACATCAGGCAGGAACCAGCACCCAAGGGC AGGCCAGCCCTCCAGCTGCCAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGAGTGCC ACAGAGGGCCCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGGCACCCCTATAACAGA ACAAATAGGTCAAGGATGCCAAATCTGAATGATTTAAAAGAGACAGCCTTGTAA28 ATGAAAATACCTAATATCGGTAACGTGATGAACAAATTTGAGATTTTGGGTGTCGTTGGCGAGGGCGCA TACGGCGTGGTTCTCAAGTGCAGACATAAGGAAACACACGAGATAGTGGCTATCAAAAAGTTTAAAGAC AGCGAAGAGAACGAGGAAGTGAAGGAGACTACTCTTCGAGAGTTGAAGATGTTGCGAACACTCAAACAA GAGAACATCGTCGAGCTTAAGGAAGCCTTTCGCAGACGCGGAAAGCTCTACTTGGTATTTGAATATGTT GAGAAAAATATGCTTGAACTTCTGGAGGAGATGCCAAATGGCGTCCCACCCGAAAAGGTTAAGAGCTAC ATCTACCAACTCATTAAAGCTATCCATTGGTGTCATAAAAACGACATCGTACACAGGGATATAAAGCCC GAGAACCTGCTTATTTCACATAATGACGTACTGAAATTGTGCGACTTCGGCTTTGCCCGGAACCTCAGC GAAGGAAATAACGCCAACTATACAGAGTATGTTGCTACTCGGTGGTACAGATCCCCAGAGCTGTTGTTG GGAGCCCCCTACGGCAAGAGTGTAGACATGTGGTCCGTCGGTTGTATCCTCGGGGAGTTGAGCGATGGG CAACCCCTGTTTCCTGGCGAGTCCGAGATCGATCAGCTGTTTACCATACAGAAAGTACTCGGACCCCTG CCATCCGAACAAATGAAATTGTTTTATTCTAATCCACGGTTCCATGGGTTGCGATTTCCTGCCGTAAAT CACCCCCAGTCACTGGAAAGGCGGTACCTCGGAATTCTGAATTCCGTACTTTTGGACCTTATGAAAAAC C T T T T GAAAC TCGACCCCGCAGACC GAT AT T T GAG C GAAC AGT GT T T GAAT C AC C C TAG AT T C C AGAC A CAGCGACTCCTTGATCGCAGCCCATCACGCTCCGCTAAGAGAAAACCTTATCACGTGGAATCTTCTACA TTGAGTAACCGAAACCAAGCAGGCAAAAGTACCGCTCTCCAAAGTCATCACCGATCAAATAGTAAGGAT ATTCAGAACCTGAGCGTAGGCTTGCCTCGAGCTGATGAGGGTCTTCCCGCCAACGAGTCTTTTCTGAAT GGAAATCTCGCAGGTGCTTCTTTGAGTCCTCTTCATACAAAGACATATCAGGCTAGCAGCCAACCAGGT TCCACTTCTAAGGATCTTACCAATAACAACATACCACATCTTCTGTCCCCAAAGGAGGCAAAGTCTAAG ACCGAGTTTGATTTCAATATAGACCCAAAGCCATCAGAAGGACCCGGTACTAAGTACCTTAAGTCAAAC TCACGCAGCCAGCAGAATCGGCACTCTTTTATGGAGAGCTCACAGTCCAAAGCCGGCACTCTCCAACCA AACGAAAAACAATCTAGGCACAGTTATATTGATACTATACCACAAAGTTCTAGGTCCCCCAGCTACAGA ACCAAGGCTAAGTCTCACGGAGCACTTAGCGATTCAAAGTCTGTAAGCAATCTCAGCGAGGCAAGGGCA CAGATTGCCGAACCCTCTACCTCCCGCTACTTTCCTTCTAGTTGCTTGGACCTCAATTCACCAACCTCA CCCACCCCCACCCGGCATAGCGACACCAGAACCCTCCTTAGCCCAAGTGGACGAAATAATCGAAACGAG GGAACCTTGGATTCACGCAGGACCACCACCCGGCATTCTAAGACCATGGAGGAACTGAAACTGCCCGAG CACATGGACAGTTCCCACTCACACTCACTGAGCGCCCCACATGAGTCCTTCTCATATGGCCTGGGATAC ACAAGTCCTTTCTCTTCCCAACAACGACCTCATAGGCACTCTATGTACGTGACCAGAGACAAGGTCAGA GCCAAAGGCTTGGATGGGTCTCTCTCCATAGGTCAGGGCATGGCAGCTCGAGCCAACAGCCTTCAGTTG CTTAGTCCTCAACCCGGTGAGCAACTGCCACCCGAAATGACCGTAGCCAGATCTTCAGTCAAAGAGACA AGCCGCGAGGGGACCTCTTCATTCCACACCCGACAGAAAAGTGAAGGTGGAGTCTATCATGACCCCCAT AGCGACGACGGTACCGCTCCTAAAGAGAATAGGCATCTGTACAACGATCCCGTGCCCCGGCGCGTTGGC AGCTTTTATCGCGTCCCTAGTCCCAGGCCAGACAACTCCTTCCATGAAAATAATGTCTCAACACGGGTT TCTAGTCTGCCCTCTGAAAGTTCATCAGGAACCAACCACAGTAAACGCCAACCCGCCTTTGACCCATGG AAAAGTCCAGAGAATATTTCCCATAGTGAACAACTGAAGGAGAAAGAGAAGCAAGGTTTCTTTAGGAGC ATGAAAAAGAAGAAAAAAAAGTCTCAGACAGTACCTAATTCTGATAGCCCTGATTTGCTCACATTGCAA AAATCCATACACTCAGCCTCCACACCTTCTAGTCGACCCAAGGAGTGGAGACCAGAGAAGATTTCCGAC CTTCAAACTCAAAGTCAACCTCTGAAGTCTCTCAGGAAACTGTTGCACCTGAGCAGCGCAAGTAACCAT CCTGCATCATCCGATCCCCGGTTCCAGCCCCTGACTGCCCAACAAACTAAAAATTCTTTCTCCGAGATA CGGATCCATCCCCTTTCCCAAGCTTCTGGTGGATCATCAAATATCCGCCAAGAACCCGCCCCAAAAGGC AGGCCCGCTCTCCAGCTTCCTGGTCAGATGGATCCTGGCTGGCACGTTAGTTCTGTAACTAGATCAGCC ACTGAAGGACCTTCATACTCAGAACAGCTTGGCGCCAAGTCCGGTCCCAATGGCCATCCTTATAACCGGACCAATAGGTCCCGCATGCCAAATCTGAACGACCTTAAGGAAACAGCTCTTTAATable 2B. Exemplary Sequences Encoding Tagged CDKL5 ProteinSEQ Nucleotide SequenceID NO.32 AT GGAGCAGAAACT CAT CT CAGAAGAGGAT CT GGGAGGT GGAGGTT CAAAGATT CCTAACATT GGTAAT GTGATGAATAAATTTGAGATCCTTGGGGTTGTAGGTGAAGGAGCCTATGGAGTTGTACTTAAATGCAGA CACAAGGAAACACAT GAAATT GT GGCGAT CAAGAAATT CAAGGACAGT GAAGAAAAT GAAGAAGT CAAAGAAACGACTTTACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAGGAAAACATTGTGGAGTTGAAGGAAAttorney Docket No. 14640.0304-00304GCATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTGAAAAAAATATGCTCGAATTGCTG GAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTAAAAAGCTACATCTATCAGCTAATCAAGGCTATT CACTGGTGCCATAAGAATGATATTGTCCATCGAGATATAAAACCAGAAAATCTCTTAATCAGCCACAAT GATGTCCTAAAACTGTGTGACTTTGGTTTTGCTCGTAATCTGTCAGAAGGCAATAATGCTAATTACACA GAGTACGTTGCCACCAGATGGTATCGGTCCCCAGAACTCTTACTTGGCGCTCCCTATGGAAAGTCCGTG GACATGTGGTCGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGACAGCCTTTATTTCCTGGAGAAAGT GAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTTCCATCTGAGCAGATGAAGCTTTTC TACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCCAGCTGTTAACCATCCTCAGTCCTTGGAAAGAAGA TACCTTGGAATTTTGAATAGTGTTCTACTTGACCTAATGAAGAATTTACTGAAGTTGGACCCAGCTGAC AGATACTTGACAGAACAGTGTTTGAATCACCCTACATTTCAAACCCAGAGACTTCTGGATCGTTCTCCT TCAAGGTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAGCCGGC AAAAGTACTGCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACATCCAGAACCTGAGTGTAGGCCTG CCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAATGGAAACCTTGCTGGAGCTAGTCTT AGTCCACTGCACACCAAAACCTACCAAGCAAGCAGCCAGCCTGGGTCTACCAGCAAAGATCTCACCAAC AAC AAC AT AC CACACCTTCTTAGCC C AAAAGAAG C C AAGT C AAAAAC AGAGT T T GAT T T T AAT AT T GAC CCAAAGCCTTCAGAAGGCCCAGGGACAAAGTACCTCAAGTCAAACAGCAGATCTCAGCAGAACCGCCAC TCATTCATGGAAAGCTCTCAAAGCAAAGCTGGGACACTGCAGCCCAATGAAAAGCAGAGTCGGCATAGC TATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGCCAAAAGCCATGGGGCA CTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCCCAAATTGCGGAGCCCAGTACCAGT AGGTACTTCCCATCTAGCTGCTTAGACTTGAATTCTCCCACCAGCCCAACCCCCACCAGACACAGTGAC ACGAGAACTTTGCTCAGCCCTTCTGGAAGAAATAACCGAAATGAGGGAACGCTGGACTCACGTCGAACC ACAACCAGACATTCTAAGACGATGGAGGAATTGAAGCTGCCGGAGCACATGGACAGTAGCCATTCCCAT TCACTGTCTGCACCTCACGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTCCCAGCAA CGTCCTCATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGAGCCAAGGGCTTGGATGGAAGCTTG AGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAACTCTTGTCACCCCAGCCTGGAGAACAG CTCCCTCCAGAGATGACTGTGGCAAGATCTTCGGTCAAAGAGACCTCCAGAGAAGGCACCTCTTCCTTC CATACACGCCAGAAGTCTGAGGGTGGAGTGTATCATGACCCACACTCTGATGATGGCACAGCCCCCAAA GAAAATAGACACCTATACAATGATCCTGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCA CGTCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAGAGAGCAGT TCTGGAACCAACCACTCAAAAAGACAACCAGCATTCGATCCATGGAAAAGTCCTGAAAATATTAGTCAT T CAGAGCAACT CAAGGAAAAAGAGAAGCAAGGATTTTT CAGGT CAAT GAAAAAGAAAAAGAAGAAAT CT CAAACAGTACCCAATTCCGACAGCCCTGATCTTCTGACGTTGCAGAAATCCATTCATTCTGCTAGCACT CCAAGCAGCAGACCAAAGGAGTGGCGCCCCGAGAAGATCTCAGATCTGCAGACCCAAAGCCAGCCATTA AAATCACTGCGCAAGTTGTTACATCTCTCTTCGGCCTCAAATCACCCGGCTTCCTCAGATCCCCGCTTC CAGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATTCGGATTCACCCCCTGAGCCAGGCC TCTGGCGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGCAGGCCAGCCCTCCAGCTGCCAGGT CAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGAGTGCCACAGAGGGCCCTTCCTACTCTGAA CAGCTGGGTGCCAAAAGTGGGCCAAATGGGCACCCCTATAACAGAACAAATCGCTCACGAATGCCAAAT C T GAAT GAT T T AAAAGAGAC AG C C T T GT AA AT GGAGCAGAAACT CAT CT CAGAAGAGGAT CT GGGAGGT GGAGGTT CAAAAATACCTAATAT CGGTAAC GTGATGAACAAATTTGAGATTTTGGGTGTCGTTGGCGAGGGCGCATACGGCGTGGTTCTCAAGTGCAGA CATAAGGAAACACACGAGATAGTGGCTATCAAAAAGTTTAAAGACAGCGAAGAGAACGAGGAAGTGAAG GAGACTACTCTTCGAGAGTTGAAGATGTTGCGAACACTCAAACAAGAGAACATCGTCGAGCTTAAGGAA GCCTTTCGCAGACGCGGAAAGCTCTACTTGGTATTTGAATATGTTGAGAAAAATATGCTTGAACTTCTG GAGGAGATGCCAAATGGCGTCCCACCCGAAAAGGTTAAGAGCTACATCTACCAACTCATTAAAGCTATC CATTGGTGTCATAAAAACGACATCGTACACAGGGATATAAAGCCCGAGAACCTGCTTATTTCACATAAT GACGTACTGAAATTGTGCGACTTCGGCTTTGCCCGGAACCTCAGCGAAGGAAATAACGCCAACTATACA GAGTATGTTGCTACTCGGTGGTACAGATCCCCAGAGCTGTTGTTGGGAGCCCCCTACGGCAAGAGTGTA GACATGTGGTCCGTCGGTTGTATCCTCGGGGAGTTGAGCGATGGGCAACCCCTGTTTCCTGGCGAGTCC GAGATCGATCAGCTGTTTACCATACAGAAAGTACTCGGACCCCTGCCATCCGAACAAATGAAATTGTTT TATTCTAATCCACGGTTCCATGGGTTGCGATTTCCTGCCGTAAATCACCCCCAGTCACTGGAAAGGCGG TACCTCGGAATTCTGAATTCCGTACTTTTGGACCTTATGAAAAACCTTTTGAAACTCGACCCCGCAGAC CGATATTTGACCGAACAGTGTTTGAATCACCCTACATTCCAGACACAGCGACTCCTTGATCGCAGCCCA TCACGCTCCGCTAAGAGAAAACCTTATCACGTGGAATCTTCTACATTGAGTAACCGAAACCAAGCAGGC AAAAGTACCGCTCTCCAAAGTCATCACCGATCAAATAGTAAGGATATTCAGAACCTGAGCGTAGGCTTG CCTCGAGCTGATGAGGGTCTTCCCGCCAACGAGTCTTTTCTGAATGGAAATCTCGCAGGTGCTTCTTTG AGTCCTCTTCATACAAAGACATATCAGGCTAGCAGCCAACCAGGTTCCACTTCTAAGGATCTTACCAAT AACAACATACCACATCTTCTGTCCCCAAAGGAGGCAAAGTCTAAGACCGAGTTTGATTTCAATATAGAC CCAAAGCCATCAGAAGGACCCGGTACTAAGTACCTTAAGTCAAACTCACGCAGCCAGCAGAATCGGCAC TCTTTTATGGAGAGCTCACAGTCCAAAGCCGGCACTCTCCAACCAAACGAAAAACAATCTAGGCACAGTTATATTGATACTATACCACAAAGTTCTAGGTCCCCCAGCTACAGAACCAAGGCTAAGTCTCACGGAGCAAttorney Docket No. 14640.0304-00304CTTAGCGATTCAAAGTCTGTAAGCAATCTCAGCGAGGCAAGGGCACAGATTGCCGAACCCTCTACCTCC CGCTACTTTCCTTCTAGTTGCTTGGACCTCAATTCACCAACCTCACCCACCCCCACCCGGCATAGCGAC ACCAGAACCCTCCTTAGCCCAAGTGGACGAAATAATCGAAACGAGGGAACCTTGGATTCACGCAGGACC ACCACCCGGCATTCTAAGACCATGGAGGAACTGAAACTGCCCGAGCACATGGACAGTTCCCACTCACAC TCACTGAGCGCCCCACATGAGTCCTTCTCATATGGCCTGGGATACACAAGTCCTTTCTCTTCCCAACAA CGACCTCATAGGCACTCTATGTACGTGACCAGAGACAAGGTCAGAGCCAAAGGCTTGGATGGGTCTCTC TCCATAGGTCAGGGCATGGCAGCTCGAGCCAACAGCCTTCAGTTGCTTAGTCCTCAACCCGGTGAGCAA CTGCCACCCGAAATGACCGTAGCCAGATCTTCAGTCAAAGAGACAAGCCGCGAGGGGACCTCTTCATTC CACACCCGACAGAAAAGTGAAGGTGGAGTCTATCATGACCCCCATAGCGACGACGGTACCGCTCCTAAA GAGAATAGGCATCTGTACAACGATCCCGTGCCCCGGCGCGTTGGCAGCTTTTATCGCGTCCCTAGTCCC AGGCCAGACAACTCCTTCCATGAAAATAATGTCTCAACACGGGTTTCTAGTCTGCCCTCTGAAAGTTCA TCAGGAACCAACCACAGTAAACGCCAACCCGCCTTTGACCCATGGAAAAGTCCAGAGAATATTTCCCAT AGT GAACAACT GAAGGAGAAAGAGAAGCAAGGTTT CTTTAGGAGCAT GAAAAAGAAGAAAAAAAAGT CT CAGACAGTACCTAATTCTGATAGCCCTGATTTGCTCACATTGCAAAAATCCATACACTCAGCCTCCACA CCTTCTAGTCGACCCAAGGAGTGGAGACCAGAGAAGATTTCCGACCTTCAAACTCAAAGTCAACCTCTG AAGTCTCTCAGGAAACTGTTGCACCTGAGCAGCGCAAGTAACCATCCTGCATCATCCGATCCCCGGTTC CAGCCCCTGACTGCCCAACAAACTAAAAATTCTTTCTCCGAGATACGGATCCATCCCCTTTCCCAAGCT TCTGGTGGATCATCAAATATCCGCCAAGAACCCGCCCCAAAAGGCAGGCCCGCTCTCCAGCTTCCTGGT CAGATGGATCCTGGCTGGCACGTTAGTTCTGTAACTAGATCAGCCACTGAAGGACCTTCATACTCAGAA CAGCTTGGCGCCAAGTCCGGTCCCAATGGCCATCCTTATAACCGGACCAATAGGTCCCGCATGCCAAATCTGAACGACCTTAAGGAAACAGCTCTTTAA2. Promoter

[0117] In some embodiments, the present disclosure provides a nucleic acid (e.g., an isolated nucleic acid) comprising a promoter operably linked to a CDKL5-encoding sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) is or is comprised within a viral genome (e.g., recombinant viral genome). Accordingly, in some embodiments, the present disclosure provides an isolated nucleic acid comprising a promoter operably linked to a CDKL5-encoding sequence and, in some embodiments, the present disclosure provides a viral genome (e.g., recombinant viral genome) comprising said isolated nucleic acid.

[0118] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5-encoding sequence and (ii) the CDKL5 -encoding sequence.

[0119] In some embodiments, the promoter is or comprises a synapsin 1 promoter. In some embodiments, the promoter is or comprises a human synapsin 1 (“human SYN1,” “hSYNl,” “hSYN,” “hSyn,” or “hSynl”) promoter.

[0120] In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto. In some embodiments, the promoter comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 18. In some embodiments, the promoter comprises a nucleotide sequence that is at least 96% identical toAttorney Docket No. 14640.0304-00304the nucleotide sequence of SEQ ID NO: 18. In some embodiments, the promoter comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of SEQ ID NO: 18. In some embodiments, the promoter comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of SEQ ID NO: 18. In some embodiments, the promoter comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 18. In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 18. In some embodiments, the promoter consists of the nucleotide sequence of SEQ ID NO: 18.

[0121] In some embodiments, the promoter is or comprises a chicken P-actin hybrid promoter (“CBh promoter”). In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 33 or a nucleotide 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 thereto. In some embodiments, the promoter comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 33. In some embodiments, the promoter comprises a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO: 33. In some embodiments, the promoter comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of SEQ ID NO: 33. In some embodiments, the promoter comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of SEQ ID NO: 33. In some embodiments, the promoter comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 33. In some embodiments, the promoter comprises the nucleotide sequence of SEQ ID NO: 33. In some embodiments, the promoter consists of the nucleotide sequence of SEQ ID NO: 33.

[0122] Non-limiting examples of promoter sequences are provided in Table 3.Table 3. Exemplary Promoter SequencesSEQ ID Nucleotide SequenceNO.18 CTGCAGAGGGCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGG GGTGCCTACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAA TTGCGCATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACT GCCAGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGC CTCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGCC ACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGAGAT AGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTCAGTCT GCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAG33 CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTAttorney Docket No. 14640.0304-00304CAATGACGGTAAATGGCCCGCCTGGCATTGTGCCCAGTACATGACCTTATGGGACTTTCCT ACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTCGAGGTGAGCCCCACGT TCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTT TTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGGGCGCGCGCCAGGCGGGGCG GGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCG GCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGCGGCGGCGGCGGCGGCCCTATAAAAAGCG AAGCGCGCGGCGGGCGGGAGTCGCTGCGCGCTGCCTTCGCCCCGTGCCCCGCTCCGCCGCC GCCTCGCGCCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAGGTGAGCGGGCGGG ACGGCCCTTCTCCTCCGGGCTGTAATTAGCTGAGCAAGAGGTAAGGGTTTAAGGGATGGTT GGTTGGTGGGGTATTAATGTTTAATTACCTGGAGCACCTGCCTGAAATCACTTTTTTTCAGGTTGG3. WPRE

[0123] In some embodiments, the present disclosure provides a nucleic acid (e.g., an isolated nucleic acid) comprising a CDKL5-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE). In some embodiments, the nucleic acid (e.g., isolated nucleic acid) is or is comprised within a viral genome (e.g., recombinant viral genome). Accordingly, in some embodiments, the present disclosure provides an isolated nucleic acid comprising a CDKL5 -encoding sequence and a WPRE and, in some embodiments, the present disclosure provides a viral genome (e.g., recombinant viral genome) comprising said isolated nucleic acid.

[0124] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, a WPRE and a CDKL5 -encoding sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, a CDKL5 -encoding sequence and a WPRE. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5 -encoding sequence, (ii) the CDKL5-encoding sequence, and (iii) a WPRE.

[0125] In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 58 or a nucleotide 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 thereto. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 58. In some embodiments, the WPRE consists of the nucleotide sequence of SEQ ID NO: 58.

[0126] In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 59 or a nucleotide 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 leastAttorney Docket No. 14640.0304-0030499%) identical thereto. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 59. In some embodiments, the WPRE consists of the nucleotide sequence of SEQ ID NO: 59.

[0127] In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 60 or a nucleotide 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 thereto. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 60. In some embodiments, the WPRE consists of the nucleotide sequence of SEQ ID NO: 60.

[0128] In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 61 or a nucleotide 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 thereto. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 61. In some embodiments, the WPRE consists of the nucleotide sequence of SEQ ID NO: 61.

[0129] In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 62 or a nucleotide 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 thereto. In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 62. In some embodiments, the WPRE consists of the nucleotide sequence of SEQ ID NO: 62.

[0130] In some embodiments, the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto. In some embodiments, the WPRE comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the WPRE comprises a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the WPRE comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the WPRE comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the WPRE comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the WPRE comprises the nucleotide sequence ofAttorney Docket No. 14640.0304-00304SEQ ID NO: 19. In some embodiments, the WPRE consists of the nucleotide sequence of SEQ ID NO: 19.

[0131] A non-limiting example of a WPRE sequence is provided in Table 4.Table 4. Exemplary WPRE SequenceSEQ ID Nucleotide SequenceNO.19 CGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTT GCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCC GTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTT GTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACT GGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTA TTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTT GGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCC TGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATC CAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCT TCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCGG58 AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGATATTCTTAACTATGTTGCTC CTTTTACGCTGTGTGGATATGCTGCTTTAATGCCTCTGTATCATGCTATTGCTTCCCGTAC GGCTTTCGTTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGG CCCGTTGTCCGTCAACGTGGCGTGGTGTGCTCTGTGTTTGCTGACGCAACCCCCACTGGCT GGGGCATTGCCACCACCTGTCAACTCCTTTCTGGGACTTTCGCTTTCCCCCTCCCGATCGC CACGGCAGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTAGGTTGCTGGGC ACTGATAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTG TTGCCAACTGGATCCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCTCTCAATCCAGC GGACCTCCCTTCCCGAGGCCTTCTGCCGGTTCTGCGGCCTCTCCCGCGTCTTCGCTTTCGG CCTCCGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTG59 AATTCGGGACATACCACGTGGTTTAGTTCCGCCTCAAACTCCAACAAATCGAGATCAAGGG AGAAAGCCTACTCCTCCAACTCCACCTCTAAGAGATACTCACCCCCACTTAACTATGAAAA ATCAGACTTTTCATCTCCAGGGGTTCGTAGACGGATTACGAGACTTGACAACAACGGAACG CCAACACAATGCCTATGGAGATCCTTTTACAACACTAAGCCCTGCGGTTCCTACTGTATCC ACCATATTGTCTCCTCCCTCGACGACTGGGGACCCTGCACTGTCACCGGAGATGTCACCAT CAAGTCTCCTAGGACTCCTCGCAGGATTACAGGTGGTGTATTTCTTGTGGACAAAAATCCT AACAATAGCTCAGAATCTAGATTGGTGGTGGACTTCTCTCAGTTTTCCAGGGGGCATACCA GAGTGCACTGGCCAAAATTCGCAGTTCCAAACTTGCAAACACTTGCCAACCTCCTGTCCAC CAACTTGCAATGGCTTTCGTTGGATGTATCTGCGGCGTTTTATCATATACCTATTAGTCCT GCTGCTGTGCCTCATCTTCTTGTTGGTTCTCCTGGACTGGAAAGGTTTAATACCTGTCTGT CCTCTTCAACCCACAACAGAAACAACAGTCAATTGCAGACAATGCACAATCTCTGCACAAG ACATGTATACTCCTCCTTACTGTTGTTGTTTAAAACCTACGGCAGGAAATTGCACTTGTTG GCCCATCCCTTCATCATGGGCTTTAGGAAATTACCTATGGGAGTGGGCCTTAGCCCGTTTC TCTTGGCTCAATTTACTAGTGCCCTTGCTTCAATGGTTAGGAGGAATTTCCCTCATTGCGT GGTTTTTGCTTATATGGATGATTTGGTTTTGGGGGCCCGCACTTCTGAGCATCTTACCGCC ATTTATTCCCATATTTGTTCTGTTTTTCTTGATTTGGGTATACATTTGAATGTCAATAAAA CAAAATGGTGGGGCAATCATCTACATTTCATGGGATATGTGATTACTAGTTCAGGTGTATT GCCACAAGACAAACATGTTAAGAAAATTTCCCGTTATTTGCACTCTGTTCCTGTTAATCAA CCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTA CGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTT CATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTT GTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCA TTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGC GGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGAC AATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTAttorney Docket No. 14640.0304-00304TCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAG ACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGTTTCGCCTCGGCGTCCGGTCCGTG TTGCTTGGTCTTCACCTGTGCAGACTTGCGAACCATGGATTCCACCGTGAACTTTGTCTCC TGGCATGCAAATCGTCAACTTGGCATGCCAAGTAAGGACCTTTGGACTCCTTATATAAAAG ATCAATTATTAACTAAATGGGAGGAGGGCAGCATTGATCCTAGATTATCAATATTTGTATT AGGAGGCTGTAGGCATAAATGCATGCGACTTCTGTAACCATGTATCTTTTTCACCTGTGCC TTGTTTTTGCCTGTGTTCCATGTCCTACTTTTCAAGCCTCCAAGCTGTGCCTTGGATGGCT TTGGGGCATGGACATAGATCCCTATAAAGAATTTGGTTCATCTTATCAGTTGTTGAATTTT CTTCCTTTGGACTTCTTTCCTGACCTTAATGCTTTGGTGGACACTGCTACTGCCTTGTATG AAGAAGAGCTAACAGGTAGGGAACATTGCTCTCCGCACCATACAGCTATTAGACAAGCTTT AGTATGCTGGGATGAATTAACTAAATTGATAGCTTGGATGAGCTCTAACATAACTTCTGAA CAAGTAAGAACAATCATAGTAAATCATGTCAATGATACCTGGGGACTTAAGGTGAGACAAA GTTTATGGTTTCATTTGTCATGTCTCACTTTCGGACAACATACAGTTCAAGAATTTTTAGT AAGTTTTGGAGTATGGATCAGAACTCCAGCTCCATATAGACCTCCTAATGCACCCATTCTC TCGACTCTTCCGGAACATACAGTCATTAGGAGAAGAGGAGGTGCAAGAGCTTCTAGGTCCC CCAGAAGACGCACTCCCTCTCCTCGCAGGAGAAGATCTCAATCACCGCGTCGCAGACGCTC TCAATCTCCATCTGCCAACTGCTGATCTTCAATGGGTACATAAAACTAATGCAATTACAGG TCTTTACTCTAACCAAGCTGCTCAGTTCAATCCGAATTGGATTCAACCTGAGTTTCCTGAA CTTCATTTACATAATGATTTAATTCAAAAATTGCAACAGTATTTTGGTCCTTTGACTATAA ATGAAAAGAGAAAATTGCAATTAAATTTTCCTGCCAGATTTTTCCCCAAAGCTACTAAATA TTTCCCTTTAATTAAAGGCATAAAAAACAATTATCCTAATTTTGCTTTAGAACATTTCTTT GCTACCGCAAATTATTTGTGGACTTTATGGGAAGCTGGAATTTTGTATTTAAGGAAGAATC AAACAACTTTGACTTTTAAAGGTAAACCATATTCTTGGGAACACAGACAGCTAGTGCAACA TAATGGGCAACAACATAAAAGTCACCTTCAATCCAGACAAAATAGCAGCATGGTGGCCTGC AGTGGGCACTTATTACACAACCACTTATCCTCAGAATCAGTCAGTGTTTCAACCAGGAATT TATCAAACAACATCTCTGATAAATCCCAAAAATCAACAAGAACTGGACTCTGTTCTTATAA ACAGATACAAACAGATAGACTGGAACACTTGGCAAGGATTTCCTGTGGATCAAAAATTTTC ATTGGTCAGCAGGGATCCTCCCCCAAAACCTTATATAAATCAATCAGCTCAAACTTTCGAA ATCAAACCTGGGCCTATAATAGTTCCCGG CGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTT GCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCC GTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTT GTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACT GGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTA TTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTT GGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCC TGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATC CAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCT TCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCG CGACTGATCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTT AACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTA TTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTA TGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCA ACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCC CCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGC TCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGG CTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGG CCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCG TCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCG CAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCT CCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTA TGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTG GCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGT TGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGAttorney Docket No. 14640.0304-00304CACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTCGCCTGT GTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAG CGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGC4. MicroRNA Binding Site

[0132] In some embodiments, the present disclosure provides a nucleic acid (e.g., an isolated nucleic acid) comprising a CDKL5-encoding sequence and a nucleotide sequence encoding at least one microRNA (miR) binding site. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) is or is comprised within a viral genome (e.g., recombinant viral genome). Accordingly, in some embodiments, the present disclosure provides an isolated nucleic acid comprising a CDKL5-encoding sequence and a nucleotide sequence encoding at least one miR binding site and, in some embodiments, the present disclosure provides a viral genome (e.g., recombinant viral genome) comprising said isolated nucleic acid.

[0133] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, a CDKL5-encoding sequence and a nucleotide sequence encoding at least one miR binding site (e.g., at least one miR183 binding site). In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5 -encoding sequence, (ii) the CDKL5 -encoding sequence, and (iii) a nucleotide sequence encoding at least one miR binding site (e.g., at least one miR183 binding site). In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5 -encoding sequence, (ii) the CDKL5 -encoding sequence, (iii) a WPRE, and (iv) a nucleotide sequence encoding at least one miR binding site (e.g., at least one miR183 binding site).

[0134] In some embodiments, the miR binding site prevents, suppresses, or otherwise inhibits expression of CDKL5 in dorsal root ganglia. In some embodiments, the miR binding site is or comprises a microRNA 183 (miR183) binding site. In some embodiments, the nucleic acid (e.g., an isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) encodes a miR binding site series comprising at least 2 miR binding sites. In some embodiments, the miR binding site series comprises at least 2, at least 3, at least 4, or at least 5 miR binding sites. In some embodiments, the miR binding site series comprises 1, 2, 3, 4, or 5 miR binding sites. In some embodiments, the miR binding sites of the miR binding site series are continuous. In some embodiments, the miR binding sites of the miR binding siteAttorney Docket No. 14640.0304-00304series are separated by a spacer. In some embodiments, the spacer is 1 to 10 nucleotides in length, e.g., 1-6 nucleotides or 5-10 nucleotides in length. In some embodiments, each miR binding site of the miR binding site series is a miR183 binding site. In some embodiments, each miR binding site of the miR binding site series is a miR183 binding site, wherein each miR183 binding site has the same nucleotide sequence.

[0135] In some embodiments, the nucleic acid (e.g., an isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence encoding at least one miR183 binding site, wherein the at least one miR183 binding site is encoded by the nucleotide sequence of AGTGAATTCTACCAGTGCCATA (SEQ ID NO: 6) or a nucleotide sequence that is at least 50% (e.g., 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%, or at least 95%) identical thereto, wherein the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of GTGCCAT. In some embodiments, the at least one miR183 binding site is encoded by a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 6 and comprises the nucleotide sequence of GTGCCAT. In some embodiments, the at least one miR183 binding site is encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 6 and comprises the nucleotide sequence of GTGCCAT. In some embodiments, the at least one miR183 binding site is encoded by a nucleotide sequence that has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, but no more than 10, modifications relative to the nucleotide sequence of SEQ ID NO: 6, wherein the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of GTGCCAT. In some embodiments, the nucleotide sequence encoding the at least one miR183 binding site has no more than 5 modifications relative to the nucleotide sequence of SEQ ID NO: 6, wherein the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of GTGCCAT. In some embodiments, the nucleotide sequence encoding the at least one miR183 binding site has 2 modifications relative to the nucleotide sequence of SEQ ID NO: 6, wherein nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of GTGCCAT. In some embodiments, the nucleotide sequence encoding the at least one miR183 binding site has 1 modification relative to the nucleotide sequence of SEQ ID NO: 6, wherein the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of GTGCCAT.

[0136] In some embodiments, the nucleotide sequence encoding the at least one miR183 binding site comprises the nucleotide sequence of SEQ ID NO: 6. In some embodiments, theAttorney Docket No. 14640.0304-00304nucleotide sequence encoding the at least one miR183 binding site consists of the nucleotide sequence of SEQ ID NO: 6.

[0137] In some embodiments, a nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) of the present disclosure comprises a nucleotide sequence encoding multiple miR183 binding sites, which may be referred to as a nucleotide sequence encoding a miR183 binding site series.

[0138] In some embodiments, the at least one miR183 binding site comprises at least two miR183 binding sites (e.g., 2, 3, 4, or 5 miR183 binding sites), wherein each miR183 binding site is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 6. In some embodiments, the miR183 binding site series comprises at least two miR183 binding sites (e.g., 2, 3, 4, or 5 miR183 binding sites), wherein each miR183 binding site is encoded by the nucleotide sequence of SEQ ID NO: 6.

[0139] In some embodiments, the miR183 binding site series comprises 4 miR binding sites. In some embodiments, the miR183 binding site series comprises 4 miR binding sites, wherein each miR183 binding site is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 6. In some embodiments, the miR183 binding site series comprises 4 miR binding sites, wherein each miR183 binding site is encoded by the nucleotide sequence of SEQ ID NO: 6.

[0140] In some embodiments, each of the miR183 binding sites in the miR183 binding site series is separated by a spacer. In some embodiments, the spacer is 1 to 10 nucleotides in length, e.g., 1-6 nucleotides or 5-10 nucleotides in length. In some embodiments, the spacer is encoded by a nucleotide sequence comprising the nucleotide sequence of GATAGTTA, or a nucleotide sequence having at least one, two, three, or four modifications, but no more than four modifications relative to the nucleotide sequence of GATAGGTA. In some embodiments, each spacer is encoded by a nucleotide sequence comprising the nucleotide sequence of GATAGTTA. In some embodiments, each spacer is encoded by the nucleotide sequence of GATAGTTA.

[0141] In some embodiments, the miR183 binding site series is encoded by the nucleotide sequence of SEQ ID NO: 7 or a nucleotide sequence that is at least 50% (e.g., 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%, or at least 95%) identical thereto, wherein at least one miR183 binding site of the series is encoded by a nucleotide sequence that comprises the nucleotide sequence of GTGCCAT. In some embodiments, the miR183 binding site series is encoded by the nucleotide sequence of SEQ ID NO: 7 or a nucleotide sequence that is at least 50% (e.g., atAttorney Docket No. 14640.0304-00304least 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%, or at least 95%) identical thereto, wherein each miR183 binding site of the series is encoded by a nucleotide sequence that comprises the nucleotide sequence of GTGCCAT. In some embodiments, the miR183 binding site series is encoded by a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 7, wherein at least one miR183 binding site of the series is encoded by a nucleotide sequence that comprises the nucleotide sequence of GTGCCAT. In some embodiments, the miR183 binding site series is encoded by a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 7, wherein each miR183 binding site of the series is encoded by a nucleotide sequence that comprises the nucleotide sequence of GTGCCAT. In some embodiments, the miR183 binding site series is encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 7, wherein at least one miR183 binding site of the series is encoded by a nucleotide sequence that comprises the nucleotide sequence of GTGCCAT. In some embodiments, the miR183 binding site series is encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 7, wherein each miR183 binding site of the series is encoded by a nucleotide sequence that comprises the nucleotide sequence of GTGCCAT. In some embodiments, the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 7. In some embodiments, the nucleotide sequence encoding the miR183 binding site series consists of the nucleotide sequence of SEQ ID NO: 7.

[0142] A non-limiting example of a nucleotide sequence encoding a miR183 binding site series is provided in Table 5.Table 5. Exemplary miR183 Binding Site-Encoding SequenceSEQ ID NO. Nucleotide Sequence7 AGTGAATTCTACCAGTGCCATAGATAGTTAAGTGAATTCTACCAGTGCCATAGATAGTTAAGTGAATTCTACCAGTGCCATAGATAGTTAAGTGAATTCTACCAGTGCCATA5. Polyadenylation Sequence

[0143] In some embodiments, the present disclosure provides a nucleic acid (e.g., an isolated nucleic acid) comprising a CDKL5-encoding sequence and a polyadenylation (poly A) sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) is or is comprised within a viral genome (e.g., recombinant viral genome). Accordingly, in some embodiments, the present disclosure provides an isolated nucleic acid comprising a CDKL5-encoding sequence and a polyA sequence and, in some embodiments, the present disclosureAttorney Docket No. 14640.0304-00304provides a viral genome (e.g., recombinant viral genome) comprising said isolated nucleic acid.

[0144] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, a CDKL5-encoding sequence and a polyA sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5 -encoding sequence, (ii) the CDKL5-encoding sequence, and (iii) a polyA sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5-encoding sequence, (ii) the CDKL5-encoding sequence, (iii) a WPRE, and (iv) a polyA sequence. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises, in 5’ to 3’ order, (i) a promoter operably linked to a CDKL5 -encoding sequence, (ii) the CDKL5-encoding sequence, (iii) a WPRE, (iv) a nucleotide sequence encoding at least one miR binding site (e.g., at least one miR183 binding site), and (v) a polyA sequence.

[0145] In some embodiments, the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto. In some embodiments, the polyA sequence comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 20. In some embodiments, the polyA sequence comprises a nucleotide sequence that is at least 96% identical to the nucleotide sequence of SEQ ID NO: 20. In some embodiments, the polyA sequence comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of SEQ ID NO: 20. In some embodiments, the polyA sequence comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of SEQ ID NO: 20. In some embodiments, the polyA sequence comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 20. In some embodiments, the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20. In some embodiments, the polyA sequence consists of the nucleotide sequence of SEQ ID NO: 20.

[0146] A non-limiting example of a polyA sequence is provided in Table 6.Table 6. Exemplary PolyA SequenceSEQ ID NO. Nucleotide SequenceAttorney Docket No. 14640.0304-0030420 CTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGAC CCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCAT TGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGA6. Inverted Terminal Repeats (ITRs)

[0147] In some embodiments, a nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) disclosed herein further comprises an ITR. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises two ITRs.

[0148] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a 5’ ITR. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a 3’ ITR. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a 5’ ITR and a 3’ ITR.

[0149] In some embodiments, the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 to the nucleotide sequence of SEQ ID NO: 17. In some embodiments, the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto. In some embodiments, the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17. In some embodiments, the 5’ ITR consists of the nucleotide sequence of SEQ ID NO: 17.

[0150] In some embodiments, the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 to the nucleotide sequence of SEQ ID NO: 21. In some embodiments, the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical thereto. In some embodiments, the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21. In some embodiments, the 3’ ITR consists of the nucleotide sequence of SEQ ID NO: 21.

[0151] Non-limiting examples of ITR sequences are provided in Table 7.Attorney Docket No. 14640.0304-00304Table 7. Exemplary ITR SequencesSEQ ID NO. Nucleotide Sequence17 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCG TCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTG GCCAACTCCATCACTAGGGGTTCCT21 AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG7. Exemplary ITR-to-ITR Sequences

[0152] In some embodiments, the present disclosure provides a nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprising a nucleotide sequence that is at least 80% (e.g., at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99%, or 100%) identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42.

[0153] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence that is at least 85% (e.g., at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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%, at least 99%, or 100%) identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42.

[0154] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide 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%, at least 99%, or 100%) identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42.

[0155] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence that is at least 95% identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence that is at least 96% identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence that is at least 97% identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42. In someAttorney Docket No. 14640.0304-00304embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence that is at least 98% identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises a nucleotide sequence that is at least 99% identical to the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42.

[0156] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) consists of the nucleotide sequence of any one of SEQ ID NOs: 22, 23, 29, 30, and 38-42.

[0157] In some embodiments, a nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) disclosed herein comprises a 5’ ITR comprising the nucleotide sequence of SEQ ID NO: 17; a promoter comprising the nucleotide sequence of SEQ ID NO: 18; a CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or 16; a WPRE comprising the nucleotide sequence of SEQ ID NO: 19; a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) further comprises a nucleotide sequence encoding a miR183 binding site series, comprising the nucleotide sequence of SEQ ID NO: 7.

[0158] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the nucleotide sequence of SEQ ID NO: 22.

[0159] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the nucleotide sequence of SEQ ID NO: 23.

[0160] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the nucleotide sequence of SEQ ID NO: 30.

[0161] In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the ITR-to-ITR sequence (the viral genome) of Construct 1. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the ITR-to-ITR sequenceAttorney Docket No. 14640.0304-00304(the viral genome) of Construct 2. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) comprises the ITR-to-ITR sequence (the viral genome) of Construct 3.

[0162] Without being bound by theory, in some embodiments, the nucleic acid (e.g., isolated nucleic acid) comprising the ITR-to-ITR components limits overexpression of CDKL5 in the presence of endogenous expression of functional CDKL5. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) comprising the ITR-to-ITR components results in a post-translational mechanism that limits overexpression of CDKL5 in the presence of endogenous expression of functional CDKL5.

[0163] Exemplary ITR-to-ITR sequences (viral genomes) are summarized in Tables 8A-8C and Table 9. In some embodiments, the nucleic acid (e.g., isolated nucleic acid) and / or viral genome (e.g., recombinant viral genome) may comprise any nucleotide sequence or combination thereof, from the nucleotide sequences in Tables 8A-8C or Table 9.Table 8A. SEQ ID NOs of Exemplary ITR-to-ITR Sequences and Components Component SEQ ID NO.Construct 1 Construct 2 Construct 35’ ITR 17 17 17Promoter 18 18 18CDKL5-encoding sequence 15 16 15WPRE 19 19 19Nucleotide sequence N / A N / A 7encoding miR183 bindingsite seriespolyA sequence 20 20 203’ ITR 21 21 21ITR-to-ITR 22 23 30Table 8B. SEQ ID NOs of Exemplary ITR-to-ITR Sequences and Components (Continued)Component SEQ ID NO.Construct 4 Construct 5 Construct 65’ ITR 17 17 17Promoter 18 18 18CDKL5-encoding sequence 15 32 43WPRE N / A N / A 19Nucleotide sequence N / A N / A N / Aencoding miR183 bindingsite seriesAttorney Docket No. 14640.0304-00304polyA sequence 20 20 203’ ITR 21 21 21ITR-to-ITR 39 42 41Table 8C. SEQ ID NOs of Exemplary ITR-to-ITR Sequences and Components (Continued)Component SEQ ID NO.Construct 7 Construct 8 Construct 95’ ITR 17 17 17Promoter 18 18 33CDKL5-encoding sequence 28 32 15WPRE 19 19 19Nucleotide sequence N / A N / A N / Aencoding miR183 bindingsite seriespolyA sequence 20 20 203’ ITR 21 21 21ITR-to-ITR 29 38 40Table 9. Exemplary Viral GenomesConstruct SEQ Nucleotide SequenceID NO.1 22 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCTGCCACCATGAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGA GATCCTTGGGGTTGTAGGTGAAGGAGCCTATGGAGTTGTACTTAAATGCAGACACAAGG AAACACAT GAAATT GT GGCGAT CAAGAAATT CAAGGACAGT GAAGAAAAT GAAGAAGT C AAAGAAACGACTTTACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAGGAAAACATTGT GGAGTTGAAGGAAGCATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTG AAAAAAATATGCTCGAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTA AAAAGCTACAT CTAT CAGCTAAT CAAGGCTATT CACT GGT GCCATAAGAAT GATATT GT CCATCGAGATATAAAACCAGAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGT GTGACTTTGGTTTTGCTCGTAATCTGTCAGAAGGCAATAATGCTAATTACACAGAGTAC GTTGCCACCAGATGGTATCGGTCCCCAGAACTCTTACTTGGCGCTCCCTATGGAAAGTC CGTGGACATGTGGTCGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGACAGCCTTTAT TTCCTGGAGAAAGTGAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTT CCATCTGAGCAGATGAAGCTTTTCTACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCC AGCTGTTAACCATCCTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTC TACTTGACCTAATGAAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAA CAGTGTTTGAATCACCCTACATTTCAAACCCAGAGACTTCTGGATCGTTCTCCTTCAAG GTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAG CCGGCAAAAGTACTGCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACATCCAGAACCTGAGTGTAGGCCTGCCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAAAttorney Docket No. 14640.0304-00304TGGAAACCTTGCTGGAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCA GCCAGCCTGGGTCTACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGC C C AAAAGAAG C C AAGT C AAAAAC AGAGT T T GAT T T T AAT AT T GAG C C AAAG C C T T C AGA AGGCCCAGGGACAAAGTACCTCAAGTCAAACAGCAGATCTCAGCAGAACCGCCACTCAT T CAT GGAAAGCT CT CAAAGCAAAGCT GGGACACT GCAGCCCAAT GAAAAGCAGAGT CGG CATAGCTATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGC CAAAAGCCATGGGGCACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGG CCCAAATTGCGGAGCCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAAT TCTCCCACCAGCCCAACCCCCACCAGACACAGTGACACGAGAACTTTGCTCAGCCCTTC TGGAAGAAATAACCGAAATGAGGGAACGCTGGACTCACGTCGAACCACAACCAGACATT CTAAGACGATGGAGGAATTGAAGCTGCCGGAGCACATGGACAGTAGCCATTCCCATTCA CTGTCTGCACCTCACGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTC CCAGCAACGTCCTCATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGAGCCAAGG GCTTGGATGGAAGCTTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAA CTCTTGTCACCCCAGCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTC GGTCAAAGAGACCTCCAGAGAAGGCACCTCTTCCTTCCATACACGCCAGAAGTCTGAGG GTGGAGTGTATCATGACCCACACTCTGATGATGGCACAGCCCCCAAAGAAAATAGACAC CTATACAATGATCCTGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCACG TCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAG AGAGCAGTTCTGGAACCAACCACTCAAAAAGACAACCAGCATTCGATCCATGGAAAAGT CCTGAAAATATTAGTCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAG GT C AAT GAAAAAGAAAAAGAAGAAAT C T C AAAC AGT AC C C AAT TCCGACAGCCCT GAT C TTCTGACGTTGCAGAAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAG TGGCGCCCCGAGAAGATCTCAGATCTGCAGACCCAAAGCCAGCCATTAAAATCACTGCG CAAGTTGTTACATCTCTCTTCGGCCTCAAATCACCCGGCTTCCTCAGATCCCCGCTTCC AGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATTCGGATTCACCCCCTG AGCCAGGCCTCTGGCGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGCAGGCC AGCCCTCCAGCTGCCAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGA GTGCCACAGAGGGCCCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGG CACCCCTATAACAGAACAAATCGCTCACGAATGCCAAATCTGAATGATTTAAAAGAGAC AGCCTTGTAAGAATTCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACT GGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTT GTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGT TGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACT GTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTC CGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTG CCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGG AAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGAC GTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGC TGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCC CTTTGGGCCGCCTCCCCGCATCGGACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGA GCTCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTC CCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATG AGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGG CAGGACAGCAAGGGGGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAG GAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGC CGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGC GAGCGCGCAGCTGCCTGCAGG23 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCTGCCACCATGAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGAGATCCTTGGGGTTGTAGGTGAAGGAGCCTATGGAGTTGTACTTAAATGCAGACACAAGGAttorney Docket No. 14640.0304-00304AAACACAT GAAATT GT GGCCAT CAAGAAATT CAAGGACAGT GAAGAAAAT GAAGAAGT C AAAGAAACCACTTTAAGGGAGCTTAAAATGCTTAGGACTCTCAAGCAGGAAAACATTGT GGAGTTGAAGGAAGCATTTAGGAGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTG AAAAGAATATGCTAGAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTA AAAAGCTACAT CTAT CAGCTAAT CAAGGCTATT CACT GGT GCCATAAGAAT GATATT GT CCATAGGGATATAAAACCAGAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGT GTGACTTTGGTTTTGCTAGGAATCTGTCAGAAGGCAATAATGCTAATTACACAGAGTAT GTTGCCACCAGATGGTATAGGTCCCCAGAACTCTTACTTGGAGCTCCCTATGGAAAGTC AGTGGACATGTGGTCTGTGGGCTGTATTCTTGGGGAGCTTAGTGATGGACAGCCTTTAT TTCCTGGAGAAAGTGAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTT CCATCTGAGCAGATGAAGCTTTTCTACAGTAATCCTAGGTTCCATGGGCTCAGGTTTCC AGCTGTTAACCATCCTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTC TACTTGACCTAATGAAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAA CAGTGTTTGAATCACCCTACATTTCAAACCCAGAGACTTCTGGATAGGTCTCCTTCAAG GTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAG CAGGCAAAAGTACT GCTTT GCAGT CT CACCACAGAT CTAACAGCAAGGACAT CCAGAAC CTGAGTGTAGGCCTGCCCAGGGCTGATGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAA TGGAAACCTTGCTGGAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCA GCCAGCCTGGGTCTACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGC C C AAAAGAAG C C AAGT C AAAAAC AGAGT T T GAT T T T AAT AT T GAC C C AAAG C C T T C AGA AGGCCCAGGGACAAAGTACCT CAAGT CAAACAGCAGAT CT CAGCAGAACAGGCACT CAT T CAT GGAAAGCT CT CAAAGCAAAGCT GGGACACT GCAGCCCAAT GAAAAGCAGAGTAGG CATAGCTATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGC CAAAAGCCATGGGGCACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGG CCCAAATTGCTGAGCCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAAT TCTCCCACCAGCCCAACCCCCACCAGACACAGTGACACCAGAACTTTGCTCAGCCCTTC TGGAAGAAATAACAGGAATGAGGGAACCCTGGACTCAAGGAGGACCACAACCAGACATT CTAAGACCATGGAGGAATTGAAGCTGCCTGAGCACATGGACAGTAGCCATTCCCATTCA CTGTCTGCACCTCATGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTC CCAGCAAAGGCCTCATAGGCATTCTATGTATGTGACCAGGGACAAAGTGAGAGCCAAGG GCTTGGATGGAAGCTTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAA CTCTTGTCACCCCAGCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTC TGTCAAAGAGACCTCCAGAGAAGGCACCTCTTCCTTCCATACAAGGCAGAAGTCTGAGG GTGGAGTGTATCATGACCCACACTCTGATGATGGCACAGCCCCCAAAGAAAATAGACAC CTATACAATGATCCTGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCAAG GCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAG AGAGCAGTTCTGGAACCAACCACTCAAAAAGACAACCAGCATTTGATCCATGGAAAAGT CCTGAAAATATTAGTCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAG GT C AAT GAAAAAGAAAAAGAAGAAAT C T C AAAC AGT AC C C AAT T C AGAC AG C C C T GAT C TTCTGACCTTGCAGAAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAG TGGAGGCCAGAGAAGATCTCAGATCTGCAGACCCAAAGCCAGCCATTAAAATCACTGAG GAAGTTGTTACATCTCTCTTCTGCCTCAAATCACCCTGCTTCCTCAGATCCCAGGTTCC AGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATTAGGATTCACCCCCTG AGCCAGGCCTCTGGAGGGAGCAGCAACATCAGGCAGGAACCAGCACCCAAGGGCAGGCC AGCCCTCCAGCTGCCAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGA GTGCCACAGAGGGCCCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGG CACCCCTATAACAGAACAAATAGGTCAAGGATGCCAAATCTGAATGATTTAAAAGAGAC AGCCTTGTAAGAATTCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACT GGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTT GTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGT TGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACT GTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTC CGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTG CCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGG AAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGAC GTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGC TGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCC CTTTGGGCCGCCTCCCCGCATCGGACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGA GCTCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTC CCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGAttorney Docket No. 14640.0304-00304CAGGACAGCAAGGGGGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAG GAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGC CGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGC GAGCGCGCAGCTGCCTGCAGG30 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCTGCCACCATGAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGA GATCCTTGGGGTTGTAGGTGAAGGAGCCTATGGAGTTGTACTTAAATGCAGACACAAGG AAACACAT GAAATT GT GGCGAT CAAGAAATT CAAGGACAGT GAAGAAAAT GAAGAAGT C AAAGAAACGACTTTACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAGGAAAACATTGT GGAGTTGAAGGAAGCATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTG AAAAAAATATGCTCGAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTA AAAAGCTACAT CTAT CAGCTAAT CAAGGCTATT CACT GGT GCCATAAGAAT GATATT GT CCATCGAGATATAAAACCAGAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGT GTGACTTTGGTTTTGCTCGTAATCTGTCAGAAGGCAATAATGCTAATTACACAGAGTAC GTTGCCACCAGATGGTATCGGTCCCCAGAACTCTTACTTGGCGCTCCCTATGGAAAGTC CGTGGACATGTGGTCGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGACAGCCTTTAT TTCCTGGAGAAAGTGAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTT CCATCTGAGCAGATGAAGCTTTTCTACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCC AGCTGTTAACCATCCTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTC TACTTGACCTAATGAAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAA CAGTGTTTGAATCACCCTACATTTCAAACCCAGAGACTTCTGGATCGTTCTCCTTCAAG GTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAG CCGGCAAAAGTACTGCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACATCCAGAAC CTGAGTGTAGGCCTGCCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAA TGGAAACCTTGCTGGAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCA GCCAGCCTGGGTCTACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGC C C AAAAGAAG C C AAGT C AAAAAC AGAGT T T GAT T T T AAT AT T GAG C C AAAG C C T T C AGA AGGCCCAGGGACAAAGTACCTCAAGTCAAACAGCAGATCTCAGCAGAACCGCCACTCAT T CAT GGAAAGCT CT CAAAGCAAAGCT GGGACACT GCAGCCCAAT GAAAAGCAGAGT CGG CATAGCTATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGC CAAAAGCCATGGGGCACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGG CCCAAATTGCGGAGCCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAAT TCTCCCACCAGCCCAACCCCCACCAGACACAGTGACACGAGAACTTTGCTCAGCCCTTC TGGAAGAAATAACCGAAATGAGGGAACGCTGGACTCACGTCGAACCACAACCAGACATT CTAAGACGATGGAGGAATTGAAGCTGCCGGAGCACATGGACAGTAGCCATTCCCATTCA CTGTCTGCACCTCACGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTC CCAGCAACGTCCTCATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGAGCCAAGG GCTTGGATGGAAGCTTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAA CTCTTGTCACCCCAGCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTC GGTCAAAGAGACCTCCAGAGAAGGCACCTCTTCCTTCCATACACGCCAGAAGTCTGAGG GTGGAGTGTATCATGACCCACACTCTGATGATGGCACAGCCCCCAAAGAAAATAGACAC CTATACAATGATCCTGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCACG TCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAG AGAGCAGTTCTGGAACCAACCACTCAAAAAGACAACCAGCATTCGATCCATGGAAAAGT CCTGAAAATATTAGTCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAG GT C AAT GAAAAAGAAAAAGAAGAAAT C T C AAAC AGT AC C C AAT TCCGACAGCCCT GAT C TTCTGACGTTGCAGAAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAG TGGCGCCCCGAGAAGATCTCAGATCTGCAGACCCAAAGCCAGCCATTAAAATCACTGCG CAAGTTGTTACATCTCTCTTCGGCCTCAAATCACCCGGCTTCCTCAGATCCCCGCTTCC AGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATTCGGATTCACCCCCTGAGCCAGGCCTCTGGCGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGCAGGCCAttorney Docket No. 14640.0304-00304AGCCCTCCAGCTGCCAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGA GTGCCACAGAGGGCCCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGG CACCCCTATAACAGAACAAATCGCTCACGAATGCCAAATCTGAATGATTTAAAAGAGAC AGCCTTGTAAGAATTCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACT GGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTT GTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGT TGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACT GTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTC CGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTG CCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGG AAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGAC GTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGC TGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCC CTTTGGGCCGCCTCCCCGCATCGGACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGA GAGT GAAT T CT AC CAGT GC CAT AGAT AGT T AAGT GAAT T CT AC CAGT GC CAT AGAT AGT TAAGTGAATTCTACCAGTGCCATAGATAGTTAAGTGAATTCTACCAGTGCCATACTCGC TGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGT GCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAA TTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGAC AGCAAGGGGGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAGGAACCC CTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCG ACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGC GCAGCTGCCTGCAGG39 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCTGCCACCATGAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGA GATCCTTGGGGTTGTAGGTGAAGGAGCCTATGGAGTTGTACTTAAATGCAGACACAAGG AAACACAT GAAATT GT GGCGAT CAAGAAATT CAAGGACAGT GAAGAAAAT GAAGAAGT C AAAGAAACGACTTTACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAGGAAAACATTGT GGAGTTGAAGGAAGCATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTG AAAAAAATATGCTCGAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTA AAAAGCTACAT CTAT CAGCTAAT CAAGGCTATT CACT GGT GCCATAAGAAT GATATT GT CCATCGAGATATAAAACCAGAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGT GTGACTTTGGTTTTGCTCGTAATCTGTCAGAAGGCAATAATGCTAATTACACAGAGTAC GTTGCCACCAGATGGTATCGGTCCCCAGAACTCTTACTTGGCGCTCCCTATGGAAAGTC CGTGGACATGTGGTCGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGACAGCCTTTAT TTCCTGGAGAAAGTGAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTT CCATCTGAGCAGATGAAGCTTTTCTACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCC AGCTGTTAACCATCCTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTC TACTTGACCTAATGAAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAA CAGTGTTTGAATCACCCTACATTTCAAACCCAGAGACTTCTGGATCGTTCTCCTTCAAG GTCAGCAAAAAGAAAACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAG CCGGCAAAAGTACTGCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACATCCAGAAC CTGAGTGTAGGCCTGCCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAA TGGAAACCTTGCTGGAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCA GCCAGCCTGGGTCTACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGC C C AAAAGAAG C C AAGT C AAAAAC AGAGT T T GAT T T T AAT AT T GAC C C AAAG C C T T C AGA AGGCCCAGGGACAAAGTACCTCAAGTCAAACAGCAGATCTCAGCAGAACCGCCACTCAT T CAT GGAAAGCT CT CAAAGCAAAGCT GGGACACT GCAGCCCAAT GAAAAGCAGAGT CGG CATAGCTATATTGACACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGC CAAAAGCCATGGGGCACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCCCAAATTGCGGAGCCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAATAttorney Docket No. 14640.0304-00304TCTCCCACCAGCCCAACCCCCACCAGACACAGTGACACGAGAACTTTGCTCAGCCCTTC TGGAAGAAATAACCGAAATGAGGGAACGCTGGACTCACGTCGAACCACAACCAGACATT CTAAGACGATGGAGGAATTGAAGCTGCCGGAGCACATGGACAGTAGCCATTCCCATTCA CTGTCTGCACCTCACGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTC CCAGCAACGTCCTCATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGAGCCAAGG GCTTGGATGGAAGCTTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAA CTCTTGTCACCCCAGCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTC GGTCAAAGAGACCTCCAGAGAAGGCACCTCTTCCTTCCATACACGCCAGAAGTCTGAGG GTGGAGTGTATCATGACCCACACTCTGATGATGGCACAGCCCCCAAAGAAAATAGACAC CTATACAATGATCCTGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCACG TCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAG AGAGCAGTTCTGGAACCAACCACTCAAAAAGACAACCAGCATTCGATCCATGGAAAAGT CCTGAAAATATTAGTCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAG GT C AAT GAAAAAGAAAAAGAAGAAAT C T C AAAC AGT AC C C AAT TCCGACAGCCCT GAT C TTCTGACGTTGCAGAAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAG TGGCGCCCCGAGAAGATCTCAGATCTGCAGACCCAAAGCCAGCCATTAAAATCACTGCG CAAGTTGTTACATCTCTCTTCGGCCTCAAATCACCCGGCTTCCTCAGATCCCCGCTTCC AGCCCTTAACAGCTCAACAAACCAAAAATTCCTTCTCAGAAATTCGGATTCACCCCCTG AGCCAGGCCTCTGGCGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGCAGGCC AGCCCTCCAGCTGCCAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGA GTGCCACAGAGGGCCCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGG CACCCCTATAACAGAACAAATCGCTCACGAATGCCAAATCTGAATGATTTAAAAGAGAC AGCCTTGTAAACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGAGCTCGCTGATCAGC CTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCT TGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCG CATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGG GGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAGGAACCCCTAGTGAT GGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGG TCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGC CTGCAGG42 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCT GCCACCAT GGAGCAGAAACT CAT CT CAGAAGAGGAT CT GGGAGGT GG AGGTTCAAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGAGATCCTTGGGGTTG TAGGT GAAGGAGCCTAT GGAGTT GTACTTAAAT GCAGACACAAGGAAACACAT GAAATT GT G G C GAT C AAGAAAT T C AAG GAC AGT GAAGAAAAT GAAGAAGT C AAAGAAAC GAC T T T ACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAGGAAAACATTGTGGAGTTGAAGGAAG CATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTGAAAAAAATATGCTC GAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTAAAAAGCTACATCTA TCAGCTAATCAAGGCTATTCACTGGTGCCATAAGAATGATATTGTCCATCGAGATATAA AACCAGAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGTGTGACTTTGGTTTT GCTCGTAATCTGTCAGAAGGCAATAATGCTAATTACACAGAGTACGTTGCCACCAGATG GTATCGGTCCCCAGAACTCTTACTTGGCGCTCCCTATGGAAAGTCCGTGGACATGTGGT CGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGACAGCCTTTATTTCCTGGAGAAAGT GAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTTCCATCTGAGCAGAT GAAGCTTTTCTACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCCAGCTGTTAACCATC CTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTCTACTTGACCTAATG AAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAACAGTGTTTGAATCA CCCTACATTTCAAACCCAGAGACTTCTGGATCGTTCTCCTTCAAGGTCAGCAAAAAGAA AACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAGCCGGCAAAAGTACT GCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACATCCAGAACCTGAGTGTAGGCCTGCCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAATGGAAACCTTGCTGAttorney Docket No. 14640.0304-00304GAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCAGCCAGCCTGGGTCT ACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGCCCAAAAGAAGCCAA GTCAAAAACAGAGTTTGATTTTAATATTGACCCAAAGCCTTCAGAAGGCCCAGGGACAA AGTACCTCAAGTCAAACAGCAGATCTCAGCAGAACCGCCACTCATTCATGGAAAGCTCT CAAAGCAAAGCTGGGACACTGCAGCCCAATGAAAAGCAGAGTCGGCATAGCTATATTGA CACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGCCAAAAGCCATGGGG CACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCCCAAATTGCGGAG CCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAATTCTCCCACCAGCCC AACCCCCACCAGACACAGTGACACGAGAACTTTGCTCAGCCCTTCTGGAAGAAATAACC GAAATGAGGGAACGCTGGACTCACGTCGAACCACAACCAGACATTCTAAGACGATGGAG GAATTGAAGCTGCCGGAGCACATGGACAGTAGCCATTCCCATTCACTGTCTGCACCTCA CGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTCCCAGCAACGTCCTC ATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGAGCCAAGGGCTTGGATGGAAGC TTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAACTCTTGTCACCCCA GCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTCGGTCAAAGAGACCT CCAGAGAAGGCACCTCTTCCTTCCATACACGCCAGAAGTCTGAGGGTGGAGTGTATCAT GACCCACACTCTGATGATGGCACAGCCCCCAAAGAAAATAGACACCTATACAATGATCC TGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCACGTCCAGACAATTCTT TCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAGAGAGCAGTTCTGGA AC C AAC C AC T C AAAAAGAC AAC C AG CAT T C GAT C CAT G GAAAAGT C C T GAAAAT AT TAG TCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAGGTCAATGAAAAAGA AAAAGAAGAAATCTCAAACAGTACCCAATTCCGACAGCCCTGATCTTCTGACGTTGCAG AAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAGTGGCGCCCCGAGAA GATCTCAGATCTGCAGACCCAAAGCCAGCCATTAAAATCACTGCGCAAGTTGTTACATC TCTCTTCGGCCTCAAATCACCCGGCTTCCTCAGATCCCCGCTTCCAGCCCTTAACAGCT CAACAAACCAAAAATTCCTTCTCAGAAATTCGGATTCACCCCCTGAGCCAGGCCTCTGG CGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGCAGGCCAGCCCTCCAGCTGC CAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGAGTGCCACAGAGGGC CCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGGCACCCCTATAACAG AACAAATCGCTCACGAATGCCAAATCTGAATGATTTAAAAGAGACAGCCTTGTAAACCC AGCTTTCTTGTACAAAGTGGGAATTCCTAGAGCTCGCTGATCAGCCTCGACTGTGCCTT CTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGT GCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAG GTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAG AGAATAGCAGGCATGCTGGGGAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTC CCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCG GGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG41 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCT GCCACCAT GGAGCAGAAACT CAT CT CAGAAGAGGAT CT GGGAGGT GG AGGTTCAAAAATACCTAATATCGGTAACGTGATGAACAAATTTGAGATTTTGGGTGTCG TTGGCGAGGGCGCATACGGCGTGGTTCTCAAGTGCAGACATAAGGAAACACACGAGATA GTGGCTATCAAAAAGTTTAAAGACAGCGAAGAGAACGAGGAAGTGAAGGAGACTACTCT TCGAGAGTTGAAGATGTTGCGAACACTCAAACAAGAGAACATCGTCGAGCTTAAGGAAG CCTTTCGCAGACGCGGAAAGCTCTACTTGGTATTTGAATATGTTGAGAAAAATATGCTT GAACTTCTGGAGGAGATGCCAAATGGCGTCCCACCCGAAAAGGTTAAGAGCTACATCTA CCAACTCATTAAAGCTATCCATTGGTGTCATAAAAACGACATCGTACACAGGGATATAA AGCCCGAGAACCTGCTTATTTCACATAATGACGTACTGAAATTGTGCGACTTCGGCTTT GCCCGGAACCTCAGCGAAGGAAATAACGCCAACTATACAGAGTATGTTGCTACTCGGTG GTACAGATCCCCAGAGCTGTTGTTGGGAGCCCCCTACGGCAAGAGTGTAGACATGTGGT CCGTCGGTTGTATCCTCGGGGAGTTGAGCGATGGGCAACCCCTGTTTCCTGGCGAGTCCGAGATCGATCAGCTGTTTACCATACAGAAAGTACTCGGACCCCTGCCATCCGAACAAATAttorney Docket No. 14640.0304-00304GAAATTGTTTTATTCTAATCCACGGTTCCATGGGTTGCGATTTCCTGCCGTAAATCACC CCCAGTCACTGGAAAGGCGGTACCTCGGAATTCTGAATTCCGTACTTTTGGACCTTATG AAAAAC C T T T T GAAAC T C GAG C C C G C AGAC C GAT AT T T GAG C GAAC AGT GT T T GAAT C A CCCTACATTCCAGACACAGCGACTCCTTGATCGCAGCCCATCACGCTCCGCTAAGAGAA AACCTTATCACGTGGAATCTTCTACATTGAGTAACCGAAACCAAGCAGGCAAAAGTACC GCTCTCCAAAGTCATCACCGATCAAATAGTAAGGATATTCAGAACCTGAGCGTAGGCTT GCCTCGAGCTGATGAGGGTCTTCCCGCCAACGAGTCTTTTCTGAATGGAAATCTCGCAG GTGCTTCTTTGAGTCCTCTTCATACAAAGACATATCAGGCTAGCAGCCAACCAGGTTCC ACTTCTAAGGATCTTACCAATAACAACATACCACATCTTCTGTCCCCAAAGGAGGCAAA GTCTAAGACCGAGTTTGATTTCAATATAGACCCAAAGCCATCAGAAGGACCCGGTACTA AGTACCTTAAGTCAAACTCACGCAGCCAGCAGAATCGGCACTCTTTTATGGAGAGCTCA CAGTCCAAAGCCGGCACTCTCCAACCAAACGAAAAACAATCTAGGCACAGTTATATTGA TACTATACCACAAAGTTCTAGGTCCCCCAGCTACAGAACCAAGGCTAAGTCTCACGGAG CACTTAGCGATTCAAAGTCTGTAAGCAATCTCAGCGAGGCAAGGGCACAGATTGCCGAA CCCTCTACCTCCCGCTACTTTCCTTCTAGTTGCTTGGACCTCAATTCACCAACCTCACC CACCCCCACCCGGCATAGCGACACCAGAACCCTCCTTAGCCCAAGTGGACGAAATAATC GAAACGAGGGAACCTTGGATTCACGCAGGACCACCACCCGGCATTCTAAGACCATGGAG GAACTGAAACTGCCCGAGCACATGGACAGTTCCCACTCACACTCACTGAGCGCCCCACA TGAGTCCTTCTCATATGGCCTGGGATACACAAGTCCTTTCTCTTCCCAACAACGACCTC ATAGGCACTCTATGTACGTGACCAGAGACAAGGTCAGAGCCAAAGGCTTGGATGGGTCT CTCTCCATAGGTCAGGGCATGGCAGCTCGAGCCAACAGCCTTCAGTTGCTTAGTCCTCA ACCCGGTGAGCAACTGCCACCCGAAATGACCGTAGCCAGATCTTCAGTCAAAGAGACAA GCCGCGAGGGGACCTCTTCATTCCACACCCGACAGAAAAGTGAAGGTGGAGTCTATCAT GACCCCCATAGCGACGACGGTACCGCTCCTAAAGAGAATAGGCATCTGTACAACGATCC CGTGCCCCGGCGCGTTGGCAGCTTTTATCGCGTCCCTAGTCCCAGGCCAGACAACTCCT TCCATGAAAATAATGTCTCAACACGGGTTTCTAGTCTGCCCTCTGAAAGTTCATCAGGA AC C AAC C AC AGT AAAC G C C AAC CCGCCTTTGACC CAT G GAAAAGT C C AGAGAAT AT T T C CCATAGTGAACAACTGAAGGAGAAAGAGAAGCAAGGTTTCTTTAGGAGCATGAAAAAGA AGAAAAAAAAGTCTCAGACAGTACCTAATTCTGATAGCCCTGATTTGCTCACATTGCAA AAATCCATACACTCAGCCTCCACACCTTCTAGTCGACCCAAGGAGTGGAGACCAGAGAA GATTTCCGACCTTCAAACTCAAAGTCAACCTCTGAAGTCTCTCAGGAAACTGTTGCACC TGAGCAGCGCAAGTAACCATCCTGCATCATCCGATCCCCGGTTCCAGCCCCTGACTGCC CAACAAACTAAAAATTCTTTCTCCGAGATACGGATCCATCCCCTTTCCCAAGCTTCTGG TGGATCATCAAATATCCGCCAAGAACCCGCCCCAAAAGGCAGGCCCGCTCTCCAGCTTC CTGGTCAGATGGATCCTGGCTGGCACGTTAGTTCTGTAACTAGATCAGCCACTGAAGGA CCTTCATACTCAGAACAGCTTGGCGCCAAGTCCGGTCCCAATGGCCATCCTTATAACCG GACCAATAGGTCCCGCATGCCAAATCTGAACGACCTTAAGGAAACAGCTCTTTAAGAAT TCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTA TGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTG CTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTAT GAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGC AACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTT TCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACA GGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTT TCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACG TCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGG CCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTC CCCGCATCGGACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGAGCTCGCTGATCAGC CTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCT TGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCG CATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGG GGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAGGAACCCCTAGTGAT GGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGG TCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGC CTGCAGG29 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCGCATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCCAttorney Docket No. 14640.0304-00304AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCTGCCACCATGAAAATACCTAATATCGGTAACGTGATGAACAAATTTGA GATTTTGGGTGTCGTTGGCGAGGGCGCATACGGCGTGGTTCTCAAGTGCAGACATAAGG AAACACACGAGATAGTGGCTATCAAAAAGTTTAAAGACAGCGAAGAGAACGAGGAAGTG AAGGAGACTACTCTTCGAGAGTTGAAGATGTTGCGAACACTCAAACAAGAGAACATCGT CGAGCTTAAGGAAGCCTTTCGCAGACGCGGAAAGCTCTACTTGGTATTTGAATATGTTG AGAAAAATATGCTTGAACTTCTGGAGGAGATGCCAAATGGCGTCCCACCCGAAAAGGTT AAGAGCTACATCTACCAACTCATTAAAGCTATCCATTGGTGTCATAAAAACGACATCGT ACACAGGGATATAAAGCCCGAGAACCTGCTTATTTCACATAATGACGTACTGAAATTGT GCGACTTCGGCTTTGCCCGGAACCTCAGCGAAGGAAATAACGCCAACTATACAGAGTAT GTTGCTACTCGGTGGTACAGATCCCCAGAGCTGTTGTTGGGAGCCCCCTACGGCAAGAG TGTAGACATGTGGTCCGTCGGTTGTATCCTCGGGGAGTTGAGCGATGGGCAACCCCTGT TTCCTGGCGAGTCCGAGATCGATCAGCTGTTTACCATACAGAAAGTACTCGGACCCCTG CCATCCGAACAAATGAAATTGTTTTATTCTAATCCACGGTTCCATGGGTTGCGATTTCC TGCCGTAAATCACCCCCAGTCACTGGAAAGGCGGTACCTCGGAATTCTGAATTCCGTAC TTTTGGACCTTATGAAAAACCTTTTGAAACTCGACCCCGCAGACCGATATTTGACCGAA CAGTGTTTGAATCACCCTACATTCCAGACACAGCGACTCCTTGATCGCAGCCCATCACG CTCCGCTAAGAGAAAACCTTATCACGTGGAATCTTCTACATTGAGTAACCGAAACCAAG CAGGCAAAAGTACCGCTCTCCAAAGTCATCACCGATCAAATAGTAAGGATATTCAGAAC CTGAGCGTAGGCTTGCCTCGAGCTGATGAGGGTCTTCCCGCCAACGAGTCTTTTCTGAA TGGAAATCTCGCAGGTGCTTCTTTGAGTCCTCTTCATACAAAGACATATCAGGCTAGCA GCCAACCAGGTTCCACTTCTAAGGATCTTACCAATAACAACATACCACATCTTCTGTCC CCAAAGGAGGCAAAGTCTAAGACCGAGTTTGATTTCAATATAGACCCAAAGCCATCAGA AGGACCCGGTACTAAGTACCTTAAGTCAAACTCACGCAGCCAGCAGAATCGGCACTCTT TTATGGAGAGCTCACAGTCCAAAGCCGGCACTCTCCAACCAAACGAAAAACAATCTAGG CACAGTTATATTGATACTATACCACAAAGTTCTAGGTCCCCCAGCTACAGAACCAAGGC TAAGTCTCACGGAGCACTTAGCGATTCAAAGTCTGTAAGCAATCTCAGCGAGGCAAGGG CACAGATTGCCGAACCCTCTACCTCCCGCTACTTTCCTTCTAGTTGCTTGGACCTCAAT TCACCAACCTCACCCACCCCCACCCGGCATAGCGACACCAGAACCCTCCTTAGCCCAAG TGGACGAAATAATCGAAACGAGGGAACCTTGGATTCACGCAGGACCACCACCCGGCATT CTAAGACCATGGAGGAACTGAAACTGCCCGAGCACATGGACAGTTCCCACTCACACTCA CTGAGCGCCCCACATGAGTCCTTCTCATATGGCCTGGGATACACAAGTCCTTTCTCTTC CCAACAACGACCTCATAGGCACTCTATGTACGTGACCAGAGACAAGGTCAGAGCCAAAG GCTTGGATGGGTCTCTCTCCATAGGTCAGGGCATGGCAGCTCGAGCCAACAGCCTTCAG TTGCTTAGTCCTCAACCCGGTGAGCAACTGCCACCCGAAATGACCGTAGCCAGATCTTC AGTCAAAGAGACAAGCCGCGAGGGGACCTCTTCATTCCACACCCGACAGAAAAGTGAAG GTGGAGTCTATCATGACCCCCATAGCGACGACGGTACCGCTCCTAAAGAGAATAGGCAT CTGTACAACGATCCCGTGCCCCGGCGCGTTGGCAGCTTTTATCGCGTCCCTAGTCCCAG GCCAGACAACTCCTTCCATGAAAATAATGTCTCAACACGGGTTTCTAGTCTGCCCTCTG AAAGTTCATCAGGAACCAACCACAGTAAACGCCAACCCGCCTTTGACCCATGGAAAAGT CCAGAGAATATTTCCCATAGTGAACAACTGAAGGAGAAAGAGAAGCAAGGTTTCTTTAG GAGCATGAAAAAGAAGAAAAAAAAGTCTCAGACAGTACCTAATTCTGATAGCCCTGATT TGCTCACATTGCAAAAATCCATACACTCAGCCTCCACACCTTCTAGTCGACCCAAGGAG TGGAGACCAGAGAAGATTTCCGACCTTCAAACTCAAAGTCAACCTCTGAAGTCTCTCAG GAAACTGTTGCACCTGAGCAGCGCAAGTAACCATCCTGCATCATCCGATCCCCGGTTCC AGCCCCTGACTGCCCAACAAACTAAAAATTCTTTCTCCGAGATACGGATCCATCCCCTT TCCCAAGCTTCTGGTGGATCATCAAATATCCGCCAAGAACCCGCCCCAAAAGGCAGGCC CGCTCTCCAGCTTCCTGGTCAGATGGATCCTGGCTGGCACGTTAGTTCTGTAACTAGAT CAGCCACTGAAGGACCTTCATACTCAGAACAGCTTGGCGCCAAGTCCGGTCCCAATGGC CATCCTTATAACCGGACCAATAGGTCCCGCATGCCAAATCTGAACGACCTTAAGGAAAC AGCTCTTTAAGAATTCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACT GGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTT GTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGT TGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACT GTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTC CGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAttorney Docket No. 14640.0304-00304AAGCTGACGTCCTTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGAC GTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGC TGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCC CTTTGGGCCGCCTCCCCGCATCGGACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGA GCTCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTC CCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATG AGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGG CAGGACAGCAAGGGGGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAG GAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGC CGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGC GAGCGCGCAGCTGCCTGCAGG38 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCTGCAGAGG GCCCTGCGTATGAGTGCAAGTGGGTTTTAGGACCAGGATGAGGCGGGGTGGGGGTGCCT ACCTGACGACCGACCCCGACCCACTGGACAAGCACCCAACCCCCATTCCCCAAATTGCG CATCCCCTATCAGAGAGGGGGAGGGGAAACAGGATGCGGCGAGGCGCGTGCGCACTGCC AGCTTCAGCACCGCGGACAGTGCCTTCGCCCCCGCCTGGCGGCGCGCGCCACCGCCGCC TCAGCACTGAAGGCGCGCTGACGTCACTCGCCGGTCCCCCGCAAACTCCCCTTCCCGGC CACCTTGGTCGCGTCCGCGCCGCCGCCGGCCCAGCCGGACCGCACCACGCGAGGCGCGA GATAGGGGGGCACGGGCGCGACCATCTGCGCTGCGGCGCCGGCGACTCAGCGCTGCCTC AGTCTGCGGTGGGCAGCGGAGGAGTCGTGTCGTGCCTGAGAGCGCAGCAAGTTTGTACA AAAAAGCAGGCT GCCACCAT GGAGCAGAAACT CAT CT CAGAAGAGGAT CT GGGAGGT GG AGGTTCAAAGATTCCTAACATTGGTAATGTGATGAATAAATTTGAGATCCTTGGGGTTG TAGGT GAAGGAGCCTAT GGAGTT GTACTTAAAT GCAGACACAAGGAAACACAT GAAATT GT G G C GAT C AAGAAAT T C AAG GAC AGT GAAGAAAAT GAAGAAGT C AAAGAAAC GAC T T T ACGAGAGCTTAAAATGCTTCGGACTCTCAAGCAGGAAAACATTGTGGAGTTGAAGGAAG CATTTCGTCGGAGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTGAAAAAAATATGCTC GAATTGCTGGAAGAAATGCCAAATGGAGTTCCACCTGAGAAAGTAAAAAGCTACATCTA TCAGCTAATCAAGGCTATTCACTGGTGCCATAAGAATGATATTGTCCATCGAGATATAA AACCAGAAAATCTCTTAATCAGCCACAATGATGTCCTAAAACTGTGTGACTTTGGTTTT GCTCGTAATCTGTCAGAAGGCAATAATGCTAATTACACAGAGTACGTTGCCACCAGATG GTATCGGTCCCCAGAACTCTTACTTGGCGCTCCCTATGGAAAGTCCGTGGACATGTGGT CGGTGGGCTGTATTCTTGGGGAGCTTAGCGATGGACAGCCTTTATTTCCTGGAGAAAGT GAAATTGACCAACTTTTTACTATTCAGAAGGTGCTAGGACCACTTCCATCTGAGCAGAT GAAGCTTTTCTACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCCAGCTGTTAACCATC CTCAGTCCTTGGAAAGAAGATACCTTGGAATTTTGAATAGTGTTCTACTTGACCTAATG AAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAACAGTGTTTGAATCA CCCTACATTTCAAACCCAGAGACTTCTGGATCGTTCTCCTTCAAGGTCAGCAAAAAGAA AACCTTACCATGTGGAAAGCAGCACATTGTCTAATAGAAACCAAGCCGGCAAAAGTACT GCTTTGCAGTCTCACCACAGATCTAACAGCAAGGACATCCAGAACCTGAGTGTAGGCCT GCCCCGGGCTGACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAATGGAAACCTTGCTG GAGCTAGTCTTAGTCCACTGCACACCAAAACCTACCAAGCAAGCAGCCAGCCTGGGTCT ACCAGCAAAGATCTCACCAACAACAACATACCACACCTTCTTAGCCCAAAAGAAGCCAA GTCAAAAACAGAGTTTGATTTTAATATTGACCCAAAGCCTTCAGAAGGCCCAGGGACAA AGTACCTCAAGTCAAACAGCAGATCTCAGCAGAACCGCCACTCATTCATGGAAAGCTCT CAAAGCAAAGCTGGGACACTGCAGCCCAATGAAAAGCAGAGTCGGCATAGCTATATTGA CACAATTCCCCAGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGCCAAAAGCCATGGGG CACTGAGTGACTCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCCCAAATTGCGGAG CCCAGTACCAGTAGGTACTTCCCATCTAGCTGCTTAGACTTGAATTCTCCCACCAGCCC AACCCCCACCAGACACAGTGACACGAGAACTTTGCTCAGCCCTTCTGGAAGAAATAACC GAAATGAGGGAACGCTGGACTCACGTCGAACCACAACCAGACATTCTAAGACGATGGAG GAATTGAAGCTGCCGGAGCACATGGACAGTAGCCATTCCCATTCACTGTCTGCACCTCA CGAATCTTTTTCTTATGGACTGGGCTACACCAGCCCCTTTTCTTCCCAGCAACGTCCTC ATAGGCATTCTATGTATGTGACCCGTGACAAAGTGAGAGCCAAGGGCTTGGATGGAAGC TTGAGCATAGGGCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAACTCTTGTCACCCCA GCCTGGAGAACAGCTCCCTCCAGAGATGACTGTGGCAAGATCTTCGGTCAAAGAGACCT CCAGAGAAGGCACCTCTTCCTTCCATACACGCCAGAAGTCTGAGGGTGGAGTGTATCAT GACCCACACTCTGATGATGGCACAGCCCCCAAAGAAAATAGACACCTATACAATGATCC TGTGCCAAGGAGAGTTGGTAGCTTTTACAGAGTGCCATCTCCACGTCCAGACAATTCTTTCCATGAAAATAATGTGTCAACTAGAGTTTCTTCTCTACCATCAGAGAGCAGTTCTGGAAttorney Docket No. 14640.0304-00304AC C AAC C AC T C AAAAAGAC AAC C AG CAT T C GAT C CAT G GAAAAGT C C T GAAAAT AT TAG TCATTCAGAGCAACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAGGTCAATGAAAAAGA AAAAGAAGAAATCTCAAACAGTACCCAATTCCGACAGCCCTGATCTTCTGACGTTGCAG AAATCCATTCATTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAGTGGCGCCCCGAGAA GATCTCAGATCTGCAGACCCAAAGCCAGCCATTAAAATCACTGCGCAAGTTGTTACATC TCTCTTCGGCCTCAAATCACCCGGCTTCCTCAGATCCCCGCTTCCAGCCCTTAACAGCT CAACAAACCAAAAATTCCTTCTCAGAAATTCGGATTCACCCCCTGAGCCAGGCCTCTGG CGGGAGCAGCAACATCCGGCAGGAACCCGCACCGAAGGGCAGGCCAGCCCTCCAGCTGC CAGGTCAGATGGATCCTGGTTGGCATGTGTCCTCTGTGACCAGGAGTGCCACAGAGGGC CCTTCCTACTCTGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGGCACCCCTATAACAG AACAAATCGCTCACGAATGCCAAATCTGAATGATTTAAAAGAGACAGCCTTGTAAGAAT TCCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTA TGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTG CTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTAT GAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGC AACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTT TCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACA GGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTT TCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACG TCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGG CCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTC CCCGCATCGGACCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGAGCTCGCTGATCAGC CTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCT TGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCG CATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGG GGAGGATTGGGAAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAGGAACCCCTAGTGAT GGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGG TCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGC CTGCAGG40 CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGT CGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGC CAACTCCATCACTAGGGGTTCCTTCTAGACAACTTTGTATAGAAAAGTTGCGTTACATA ACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAA TAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACT GCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAA TGACGGTAAATGGCCCGCCTGGCATTGTGCCCAGTACATGACCTTATGGGACTTTCCTA CTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTCGAGGTGAGCCCCACG TTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTAT TTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGGGGCGCGCGCCAGGCG GGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAAT CAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGCGGCGGCGGCGGCGGCCCTA TAAAAAGCGAAGCGCGCGGCGGGCGGGAGTCGCTGCGCGCTGCCTTCGCCCCGTGCCCC GCTCCGCCGCCGCCTCGCGCCGCCCGCCCCGGCTCTGACTGACCGCGTTACTCCCACAG GTGAGCGGGCGGGACGGCCCTTCTCCTCCGGGCTGTAATTAGCTGAGCAAGAGGTAAGG GTTTAAGGGATGGTTGGTTGGTGGGGTATTAATGTTTAATTACCTGGAGCACCTGCCTG AAATCACTTTTTTTCAGGTTGGCAAGTTTGTACAAAAAAGCAGGCTGCCACCATGAAGA TTCCTAACATTGGTAATGTGATGAATAAATTTGAGATCCTTGGGGTTGTAGGTGAAGGA GCCTATGGAGTTGTACTTAAATGCAGACACAAGGAAACACATGAAATTGTGGCGATCAA GAAAT T C AAG GAC AGT GAAGAAAAT GAAGAAGT C AAAGAAAC GACTTTACGAGAGCTTA AAATGCTTCGGACTCTCAAGCAGGAAAACATTGTGGAGTTGAAGGAAGCATTTCGTCGG AGGGGAAAGTTGTACTTGGTGTTTGAGTATGTTGAAAAAAATATGCTCGAATTGCTGGA AGAAAT GCCAAAT GGAGTT CCACCT GAGAAAGTAAAAAGCTACAT CTAT CAGCTAAT CA AGGCTATTCACTGGTGCCATAAGAATGATATTGTCCATCGAGATATAAAACCAGAAAAT CTCTTAATCAGCCACAATGATGTCCTAAAACTGTGTGACTTTGGTTTTGCTCGTAATCT GTCAGAAGGCAATAATGCTAATTACACAGAGTACGTTGCCACCAGATGGTATCGGTCCC CAGAACTCTTACTTGGCGCTCCCTATGGAAAGTCCGTGGACATGTGGTCGGTGGGCTGT ATTCTTGGGGAGCTTAGCGATGGACAGCCTTTATTTCCTGGAGAAAGTGAAATTGACCA ACTTTTTACTATTCAGAAGGTGCTAGGACCACTTCCATCTGAGCAGATGAAGCTTTTCT ACAGTAATCCTCGCTTCCATGGGCTCCGGTTTCCAGCTGTTAACCATCCTCAGTCCTTG GAAAGAAGATACCTTGGAATTTTGAATAGTGTTCTACTTGACCTAATGAAGAATTTACTGAAGTTGGACCCAGCTGACAGATACTTGACAGAACAGTGTTTGAATCACCCTACATTTCAttorney Docket No. 14640.0304-00304AAACCCAGAGACTTCTGGATCGTTCTCCTTCAAGGTCAGCAAAAAGAAAACCTTACCAT GTGGAAAGCAGCACATTGTCTAATAGAAACCAAGCCGGCAAAAGTACTGCTTTGCAGTC TCACCACAGATCTAACAGCAAGGACATCCAGAACCTGAGTGTAGGCCTGCCCCGGGCTG ACGAAGGTCTCCCTGCCAATGAAAGCTTCCTAAATGGAAACCTTGCTGGAGCTAGTCTT AGTCCACTGCACACCAAAACCTACCAAGCAAGCAGCCAGCCTGGGTCTACCAGCAAAGA TCTCACCAACAACAACATACCACACCTTCTTAGCCCAAAAGAAGCCAAGTCAAAAACAG AGTTTGATTTTAATATTGACCCAAAGCCTTCAGAAGGCCCAGGGACAAAGTACCTCAAG T CAAACAGCAGAT CT CAGCAGAACCGCCACT GATT CAT GGAAAGCT CT CAAAGCAAAGC TGGGACACTGCAGCCCAATGAAAAGCAGAGTCGGCATAGCTATATTGACACAATTCCCC AGTCCTCTAGGAGTCCCTCCTACAGGACCAAGGCCAAAAGCCATGGGGCACTGAGTGAC TCCAAGTCTGTGAGCAACCTTTCTGAAGCCAGGGCCCAAATTGCGGAGCCCAGTACCAG TAGGTACTTCCCATCTAGCTGCTTAGACTTGAATTCTCCCACCAGCCCAACCCCCACCA GACACAGTGACACGAGAACTTTGCTCAGCCCTTCTGGAAGAAATAACCGAAATGAGGGA ACGCTGGACTCACGTCGAACCACAACCAGACATTCTAAGACGATGGAGGAATTGAAGCT GCCGGAGCACATGGACAGTAGCCATTCCCATTCACTGTCTGCACCTCACGAATCTTTTT CTTATGGACTGGGCTACACCAGCCCCTTTTCTTCCCAGCAACGTCCTCATAGGCATTCT ATGTATGTGACCCGTGACAAAGTGAGAGCCAAGGGCTTGGATGGAAGCTTGAGCATAGG GCAAGGGATGGCAGCTAGAGCCAACAGCCTGCAACTCTTGTCACCCCAGCCTGGAGAAC AGCTCCCTCCAGAGATGACTGTGGCAAGATCTTCGGTCAAAGAGACCTCCAGAGAAGGC ACCTCTTCCTTCCATACACGCCAGAAGTCTGAGGGTGGAGTGTATCATGACCCACACTC TGATGATGGCACAGCCCCCAAAGAAAATAGACACCTATACAATGATCCTGTGCCAAGGA GAGTTGGTAGCTTTTACAGAGTGCCATCTCCACGTCCAGACAATTCTTTCCATGAAAAT AATGTGTCAACTAGAGTTTCTTCTCTACCATCAGAGAGCAGTTCTGGAACCAACCACTC AAAAAGAC AAC C AG CAT T C GAT C CAT G GAAAAGT C C T GAAAAT AT T AGT CAT T C AGAG C AACTCAAGGAAAAAGAGAAGCAAGGATTTTTCAGGTCAATGAAAAAGAAAAAGAAGAAA TCTCAAACAGTACCCAATTCCGACAGCCCTGATCTTCTGACGTTGCAGAAATCCATTCA TTCTGCTAGCACTCCAAGCAGCAGACCAAAGGAGTGGCGCCCCGAGAAGATCTCAGATC TGCAGACCCAAAGCCAGCCATTAAAATCACTGCGCAAGTTGTTACATCTCTCTTCGGCC TCAAATCACCCGGCTTCCTCAGATCCCCGCTTCCAGCCCTTAACAGCTCAACAAACCAA AAATTCCTTCTCAGAAATTCGGATTCACCCCCTGAGCCAGGCCTCTGGCGGGAGCAGCA ACATCCGGCAGGAACCCGCACCGAAGGGCAGGCCAGCCCTCCAGCTGCCAGGTCAGATG GATCCTGGTTGGCATGTGTCCTCTGTGACCAGGAGTGCCACAGAGGGCCCTTCCTACTC TGAACAGCTGGGTGCCAAAAGTGGGCCAAATGGGCACCCCTATAACAGAACAAATCGCT C AC GAAT G C C AAAT C T GAAT GAT T T AAAAGAGAC AG C C T T GT AAGAAT T C C GAT AAT C A ACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTT TTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATG GCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTG GCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTG GTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCT ATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCT GTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGC TCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCC CTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCG TCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCGGA CCCAGCTTTCTTGTACAAAGTGGGAATTCCTAGAGCTCGCTGATCAGCCTCGACTGTGC CTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAA GGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAG TAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGG AAGAGAATAGCAGGCATGCTGGGGAGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCA CTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG B. Associated Virus (AAV) Capsids and Particles

[0164] AAVs typically have a genome of about 5,000 nucleotides in length and contain one open reading frame encoding the (non-structural) proteins responsible for replication (Rep78, Rep68, Rep52, Rep40, encoded by Rep genes) and another open reading frame encoding the structural proteins of the capsid (VP1, VP2, VP3, encoded by capsid genes or Cap genes).Attorney Docket No. 14640.0304-00304The open reading frames are flanked by two inverted terminal repeat (ITR) sequences, which serve as the origin of replication of the viral genome. 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. VP1 is the full-length capsid protein sequence and contains the VP2 and VP3 sequences and VP2 and VP3 are shorter components of the whole, with the VP2 sequence containing the VP3 sequence. Though it varies by AAV serotype, as a non-limiting example, for AAV9 / hu.14 (SEQ ID NO: 123 of US 7,906,111, the relevant 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. With reference to the AAV9 capsid variant TTM-027, TTM-003, or TTM-043, VP1 comprises amino acids 1-742, VP2 comprises amino acids 138-742, and VP3 comprises amino acids 203-742.

[0165] Changes in the sequence in the VP3 region of a single capsid open reading frame 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 the 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.

[0166] 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.

[0167] AAV particles may be used as a biological tool, including in gene therapy, 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. Moreover, infection with AAV particles has minimal influence on changing the pattern of cellular gene expression (Stilwell and Samulski et al., Biotechniques, 2003, 34, 148, the relevant contents of which are herein incorporated by reference in their entirety). 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 payload.Attorney Docket No. 14640.0304-00304

[0168] Typically, AAV particles for CDKL5 delivery may be recombinant viral particles that 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 CDKL5. In some cases, the nucleic acid sequence encoding CDKL5 further comprises one or more regulatory elements to modulate transcription.

[0169] In some embodiments, the AAV particles of the present disclosure may be introduced into mammalian cells.

[0170] 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 (scAAV) viral genomes. scAAV viral genomes contain DNA strands that anneal together to form doublestranded 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.

[0171] Methods for producing and / or modifying AAV particles are disclosed in the art such as pseudotyped AAV particles (International Patent Publication Nos. W0200028004;W0200123001; WO2004112727; W02005005610; and W02005072364, the relevant contents of each of which are incorporated herein by reference in their entirety).

[0172] In some embodiments, an AAV particle of the present disclosure comprises a viral genome (e.g., recombinant viral genome) disclosed herein and an AAV capsid. In some embodiments, the AAV capsid is an AAV capsid variant, wherein the variant comprises a capsid protein that differs from a wildtype AAV capsid protein by one or more insertions, deletions, and / or substitutions. In some embodiments, the AAV capsid or AAV capsid variant comprises a capsid protein selected from the group consisting of: AAV1, AAV2, AAV3, AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAV9, AAV9 K449R, VOY101, VOY201, AAVPHP. A, AAVPHP. B, AAVPHP. B2, AAVPHP. B3, AAVPHP.eB, AAVPHP. N, AAVPHP. S, G2B4, G2B5, CAP-B10, AAVrhlO, AAVrh32.33, AAVrh74, a capsid protein of an AAV serotype as provided in Table 6 of International Patent Publication No.WO2021230987 (the relevant contents of which are incorporated by reference in their entirety), a capsid protein disclosed in International Patent Publication No. WO2023081648 (the relevant contents of which are incorporated by reference in their entirety), a capsid protein disclosed in International Patent Publication No. WO2023154693 (the relevant contents of which are incorporated by reference in their entirety), a capsid protein disclosedAttorney Docket No. 14640.0304-00304in International Patent Publication No. WO2023235791 (the relevant contents of which are incorporated by reference in their entirety), and a capsid protein disclosed in International Patent Publication No. W02024006741 (the relevant contents of which are incorporated by reference in their entirety), or a variant of any of the foregoing.

[0173] In some embodiments, the AAV capsid variant preferentially targets the brain over the liver. In some embodiments, the AAV capsid variant is AAVPHP.eB or CAP -B 10. See, e.g., Goertsen et al. AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset. Nat Neurosci 25, 106-115 (2022); Seo et al. Multimodal imaging of capsid and cargo reveals differential brain targeting and liver detargeting of systemically-administered AAVs, Biomaterials 288 (2022). In some embodiments, the AAV capsid variant is an AAV9 capsid variant disclosed in International Patent Publication No. WO2023081648 or WO2023235791. In some embodiments, the AAV5 capsid variant is an AAV9 capsid variant disclosed in International Patent Publication No. WO2023154693.

[0174] In some embodiments, the present disclosure provides an AAV particle comprising a viral genome (e.g., recombinant viral genome) disclosed herein and an AAV9 capsid variant. In some embodiments, the AAV9 capsid variant comprises at least one insertion and / or at least one substitution in hypervariable loop IV relative to a wildtype AAV9 capsid. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop IV is present in a VP1 of the AAV9 capsid variant. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop IV is present in a VP2 of the AAV9 capsid variant. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop IV is present in a VP3 of the AAV9 capsid variant. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop IV is present in a VP1, VP2, and VP3 of the AAV9 capsid variant.

[0175] In some embodiments, the AAV9 capsid variant comprises at least one insertion and / or at least one substitution in hypervariable loop VIII relative to a wildtype AAV9 capsid. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop VIII is present in a VP1 of the AAV9 capsid variant. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop VIII is present in a VP2 of the AAV9 capsid variant. In some embodiments, the at least one insertion and / or at least one substitution in hypervariable loop VIII is present in a VP3 of the AAV9 capsid variant. In some embodiments, the at least one insertion and / or at least oneAttorney Docket No. 14640.0304-00304substitution in hypervariable loop VIII is present in a VP1, VP2, and VP3 of the AAV9 capsid variant.

[0176] In some embodiments, the at least one insertion and / or at least one substitution increases 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, the cortex (e.g., frontal, parietal, occipital, and / or temporal), thalamus, hypothalamus, striatum, putamen, caudate nucleus, hippocampus, entorhinal cortex, basal ganglia, or deep cerebellar nuclei.

[0177] In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the cortex (e.g., the motor cortex). In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the hippocampus. In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the striatum. In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the thalamus. In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the cerebellum. In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to a glutamatergic neuron. In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to a GABAergic neuron.

[0178] In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the CNS (e.g., the cortex) after intravenous administration. In some embodiments, the at least one insertion and / or at least one substitution increases distribution of an AAV particle to the CNS (e.g., the cortex) following focused ultrasound (FUS), e.g., coupled with the intravenous administration of microbubbles (FUS-MB), or MRI-guided FUS coupled with intravenous administration.

[0179] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of ENVSGSPHSKA (SEQ ID NO: 8) in hypervariable loop IV. In some embodiments, the amino acid sequence of SEQ ID NO: 8 is present in VP1, VP2, and / or VP3. In some embodiments, the amino acid sequence of SEQ ID NO: 8 is present in VP1, VP2, and VP3.Attorney Docket No. 14640.0304-00304

[0180] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 5) in hypervariable loop IV. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 5 present at positions 254-259 numbered according to the amino acid sequence of SEQ ID NO: 25, comprises the amino acid E at position 249 numbered according to the amino acid sequence of SEQ ID NO: 25, and comprises the amino acid V at position 251 numbered according to the amino acid sequence of SEQ ID NO: 25. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 5 present at positions 319-324 numbered according to the amino acid sequence of SEQ ID NO: 24, comprises the amino acid E at position 314 numbered according to the amino acid sequence of SEQ ID NO: 24, and comprises the amino acid V at position 316 numbered according to the amino acid sequence of SEQ ID NO: 24. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 5 present at positions 456-461 numbered according to the amino acid sequence of SEQ ID NO: 1, comprises the amino acid E at position 451 numbered according to the amino acid sequence of SEQ ID NO: 1, and comprises the amino acid V at position 453 numbered according to the amino acid sequence of SEQ ID NO: 1.

[0181] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequenceAttorney Docket No. 14640.0304-00304that is at least 97% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, the amino acid sequence of SEQ ID NO: 25 may be referred to as VP3 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a VP3 comprising the amino acid sequence of SEQ ID NO: 25. In some embodiments, an AAV particle of the present disclosure comprises a VP3 consisting of the amino acid sequence of SEQ ID NO: 25.

[0182] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 24 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 314-324 of the amino acid sequence ofAttorney Docket No. 14640.0304-00304SEQ ID NO: 24 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, the amino acid sequence of SEQ ID NO: 24 may be referred to as VP2 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a VP2 comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, an AAV particle of the present disclosure comprises a VP2 consisting of the amino acid sequence of SEQ ID NO: 24.

[0183] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 1 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 1, whereinAttorney Docket No. 14640.0304-00304the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 8 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the amino acid sequence of SEQ ID NO: 1 may be referred to as VP1 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a VP1 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, an AAV particle of the present disclosure comprises a VP1 consisting of the amino acid sequence of SEQ ID NO: 1.

[0184] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) in hypervariable loop IV. In some embodiments, the amino acid sequence of SEQ ID NO: 9 is present in VP1, VP2, and / or VP3. In some embodiments, the amino acid sequence of SEQ ID NO: 9 is present in VP1, VP2, and VP3.

[0185] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 247-264 ofAttorney Docket No. 14640.0304-00304the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 25, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 25). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, the amino acid sequence of SEQ ID NO: 25 may be referred to as VP3 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a VP3 comprising the amino acid sequence of SEQ ID NO: 25. In some embodiments, an AAV particle of the present disclosure comprises a VP3 consisting of the amino acid sequence of SEQ ID NO: 25.

[0186] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 24 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 thereto, wherein the AAV9 capsid variant comprises theAttorney Docket No. 14640.0304-00304amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 312-329 of an amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 312-329 of an amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises an amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises an amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 24, wherein the AAV9 capsid variant comprises an amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 24). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, the amino acid sequence of SEQ ID NO: 24 may be referred to as VP2 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a VP2 comprising the amino acid sequence of SEQ ID NO: 24. In some embodiments, an AAV particle of the present disclosure comprises a VP2 consisting of the amino acid sequence of SEQ ID NO: 24.

[0187] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence that is at least 90% (e.g., at least 90%, at leastAttorney Docket No. 14640.0304-0030491%, 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 AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 1, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 9 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 1). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the amino acid sequence of SEQ ID NO: 1 may be referred to as VP1 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a VP1 comprising the amino acid sequence of SEQ ID NO: 1. In some embodiments, an AAV particle of the present disclosure comprises a VP1 consisting of the amino acid sequence of SEQ ID NO: 1.Attorney Docket No. 14640.0304-00304

[0188] In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant that is encoded by the nucleotide sequence of SEQ ID NO: 2 or a nucleotide sequence that is 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%) identical thereto. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 80% identical to the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 2. In some embodiments, the AAV capsid variant is encoded by the nucleotide sequence of SEQ ID NO: 2.

[0189] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of ERVSGSPHSKA (SEQ ID NO: 10) in hypervariable loop IV. In some embodiments, the amino acid sequence of SEQ ID NO: 10 is present in VP1, VP2, and / or VP3. In some embodiments, the amino acid sequence of SEQ ID NO: 10 is present in VP1, VP2, and VP3.

[0190] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 5) in hypervariable loop IV. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 5 present at positions 254-259 numbered according to the amino acid sequence of SEQ ID NO: 27, comprises the amino acid E at position 249 numbered according to the amino acid sequence of SEQ ID NO: 27, comprises the amino acid R at position 250 numbered according to the amino acid sequence of SEQ ID NO: 27, and comprises the amino acid V at position 251 numbered according to the amino acid sequence of SEQ ID NO: 27. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 5 present at positions 319-324 numbered according to the amino acid sequence of SEQ ID NO: 26, comprises the amino acid E at position 314 numbered according to the amino acid sequence of SEQ ID NO: 26, comprises the amino acid R at position 315 numbered according to the amino acid sequence of SEQ ID NO: 26, and comprises the amino acid V at position 316 numbered according to the amino acid sequence of SEQ ID NO: 26. In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 5 present at positions 456-461 numbered according toAttorney Docket No. 14640.0304-00304the amino acid sequence of SEQ ID NO: 4, comprises the amino acid E at position 451 numbered according to the amino acid sequence of SEQ ID NO: 4, comprises the amino acid R at position 452 numbered according to the amino acid sequence of SEQ ID NO: 4, and comprises the amino acid V at position 453 numbered according to the amino acid sequence of SEQ ID NO: 4.

[0191] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 249-259 of theAttorney Docket No. 14640.0304-00304amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 249-259 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the amino acid sequence of SEQ ID NO: 27 may be referred to as VP3 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a VP3 comprising the amino acid sequence of SEQ ID NO: 27. In some embodiments, an AAV particle of the present disclosure comprises a VP3 consisting of the amino acid sequence of SEQ ID NO: 27.

[0192] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 26 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the aminoAttorney Docket No. 14640.0304-00304acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 314-324 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the amino acid sequence of SEQ ID NO: 26 may be referred to as VP2 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a VP2 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, an AAV particle of the present disclosure comprises a VP2 consisting of the amino acid sequence of SEQ ID NO: 26.

[0193] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 4 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 451-461 of the amino acidAttorney Docket No. 14640.0304-00304sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 10 present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 451-461 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the amino acid sequence of SEQ ID NO: 4 may be referred to as VP1 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a VP1 comprising the amino acid sequence of SEQ ID NO: 4. In some embodiments, an AAV particle of the present disclosure comprises a VP1 consisting of the amino acid sequence of SEQ ID NO: 4.

[0194] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) in hypervariable loop IV. In some embodiments, the amino acid sequence of SEQ ID NO: 11 is present in VP1, VP2, and / or VP3. In some embodiments, the amino acid sequence of SEQ ID NO: 11 is present in VP1, VP2, and VP3.

[0195] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 27, wherein theAttorney Docket No. 14640.0304-00304AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 27, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 27). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the amino acid sequence of SEQ ID NO: 27 may be referred to as VP3 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a VP3 comprising the amino acid sequence of SEQ ID NO: 27. In some embodiments, an AAV particle of the present disclosure comprises a VP3 consisting of the amino acid sequence of SEQ ID NO: 27.

[0196] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 26 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 312-329 of the amino acid sequence of SEQ IDAttorney Docket No. 14640.0304-00304NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 26, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 26). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 26. In some embodiments, the amino acid sequence of SEQ ID NO: 26 may be referred to as VP2 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a VP2 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, an AAV particle of the present disclosure comprises a VP2 consisting of the amino acid sequence of SEQ ID NO: 26.

[0197] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 4 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present atAttorney Docket No. 14640.0304-00304amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 4, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 11 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 4). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 4. In some embodiments, the amino acid sequence of SEQ ID NO: 4 may be referred to as VP1 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a VP1 comprising the amino acid sequence of SEQ ID NO: 4. In some embodiments, an AAV particle of the present disclosure comprises a VP1 consisting of the amino acid sequence of SEQ ID NO: 4.

[0198] In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant that is encoded by the nucleotide sequence of SEQ ID NO: 3 or a nucleotide sequence that is 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%) identical thereto. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 80% identical to the nucleotide sequence of SEQ ID NO: 3. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 3. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 3. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 99% identical to the nucleotide sequence ofAttorney Docket No. 14640.0304-00304SEQ ID NO: 3. In some embodiments, the AAV capsid variant is encoded by the nucleotide sequence of SEQ ID NO: 3.

[0199] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV and the amino acid sequence of RQTALQL (SEQ ID NO: 49) in hypervariable loop VIII. In some embodiments, the amino acid sequences of SEQ ID NO: 12 and SEQ ID NO: 49 are present in VP1, VP2, and / or VP3. In some embodiments, the amino acid sequences of SEQ ID NO: 12 and SEQ ID NO: 49 are present in VP1, VP2, and VP3.

[0200] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV (e.g., present at positions 252-257 numbered according to the amino acid sequence of SEQ ID NO: 47) and three, four, or all of the amino acid R at position 388, the amino acid T at position 390, the amino acid L at position 392, the amino acid Q at position 393, and / or the amino acid L at position 394, each numbered according to the amino acid sequence of SEQ ID NO: 47. In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV (e.g., present at positions 252-257 numbered according to the amino acid sequence of SEQ ID NO: 47) and the amino acid R at position 388, the amino acid T at position 390, the amino acid L at position 392, the amino acid Q at position 393, and the amino acid L at position 394, each numbered according to the amino acid sequence of SEQ ID NO: 47.

[0201] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV (e.g., present at positions 317-322 numbered according to the amino acid sequence of SEQ ID NO: 46) and three, four, or all of the amino acid R at position 453, the amino acid T at position 455, the amino acid L at position 457, the amino acid Q at position 458, and / or the amino acid L at position 459, each numbered according to the amino acid sequence of SEQ ID NO: 46. In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV (e.g., present at positions 317-322 numbered according to the amino acid sequence of SEQ ID NO: 46) and the amino acid R at position 453, the amino acid T at position 455, the amino acid L at position 457, the amino acid Q at position 458, and the amino acid L at position 459, each numbered according to the amino acid sequence of SEQ ID NO: 46.Attorney Docket No. 14640.0304-00304

[0202] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV (e.g., present at positions 454-459 numbered according to the amino acid sequence of SEQ ID NO: 45) and three, four, or all of the amino acid R at position 590, the amino acid T at position 592, the amino acid L at position 594, the amino acid Q at position 595, and / or the amino acid L at position 596, each numbered according to the amino acid sequence of SEQ ID NO: 45. In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of HDSPHK (SEQ ID NO: 12) in hypervariable loop IV (e.g., present at positions 454-459 numbered according to the amino acid sequence of SEQ ID NO: 45) and the amino acid R at position 590, the amino acid T at position 592, the amino acid L at position 594, the amino acid Q at position 595, and the amino acid L at position 596, each numbered according to the amino acid sequence of SEQ ID NO: 45.

[0203] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 252-257 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 252-257 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positionsAttorney Docket No. 14640.0304-00304252-257 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 252-257 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 252-257 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 252-257 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47. In some embodiments, the amino acid sequence of SEQ ID NO: 47 may be referred to as VP3 of TTM-043. In some embodiments, an AAV particle of the present disclosure comprises a VP3 comprising the amino acid sequence of SEQ ID NO: 47. In some embodiments, an AAV particle of the present disclosure comprises a VP3 consisting of the amino acid sequence of SEQ ID NO: 47.

[0204] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 46 or an amino acid sequence that is at least 90% (e.g., at least 90%, at leastAttorney Docket No. 14640.0304-0030491%, 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 AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 317-322 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 317-322 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 317-322 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 317-322 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positionsAttorney Docket No. 14640.0304-00304317-322 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 317-322 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 46. In some embodiments, the amino acid sequence of SEQ ID NO: 46 may be referred to as VP2 of TTM-043. In some embodiments, an AAV particle of the present disclosure comprises a VP2 comprising the amino acid sequence of SEQ ID NO: 46. In some embodiments, an AAV particle of the present disclosure comprises a VP2 consisting of the amino acid sequence of SEQ ID NO: 46.

[0205] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 45 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 454-459 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 454-459 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In someAttorney Docket No. 14640.0304-00304embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 454-459 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 454-459 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 454-459 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 12 present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 454-459 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of SEQ ID NO: 49 present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 45. In some embodiments, the amino acid sequence of SEQ ID NO: 45 may be referred to as VP1 of TTM-043. In some embodiments, an AAV particle of the present disclosure comprises a VP1 comprising the amino acid sequence of SEQ ID NO: 45. In some embodiments, anAttorney Docket No. 14640.0304-00304AAV particle of the present disclosure comprises a VP1 consisting of the amino acid sequence of SEQ ID NO: 45.

[0206] In some embodiments, an AAV particle of the present disclosure comprises an AAV9 capsid variant comprising the amino acid sequence of KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) in hypervariable loop IV and the amino acid sequence of RQTALQL (SEQ ID NO: 49) in hypervariable loop VIII. In some embodiments, the amino acid sequences of SEQ ID NO: 13 and RQTALQL (SEQ ID NO: 49) are present in VP1, VP2, and / or VP3. In some embodiments, the amino acid sequences of SEQ ID NO: 13 and RQTALQL (SEQ ID NO: 49) are present in VP1, VP2, and VP3.

[0207] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In someAttorney Docket No. 14640.0304-00304embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 47, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 247-264 of the amino acid sequence of SEQ ID NO: 47) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47 (e.g., present at positions 388-394 of the amino acid sequence of SEQ ID NO: 47). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47. In some embodiments, the amino acid sequence of SEQ ID NO: 47 may be referred to as VP3 of TTM-043. In some embodiments, an AAV particle of the present disclosure comprises a VP3 comprising the amino acid sequence of SEQ ID NO: 47. In some embodiments, an AAV particle of the present disclosure comprises a VP3 consisting of the amino acid sequence of SEQ ID NO: 47.

[0208] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 46 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positionsAttorney Docket No. 14640.0304-00304312-329 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence ofAttorney Docket No. 14640.0304-00304SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 46, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 312-329 of the amino acid sequence of SEQ ID NO: 46) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46 (e.g., present at positions 453-459 of the amino acid sequence of SEQ ID NO: 46). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 46. In some embodiments, the amino acid sequence of SEQ ID NO: 46 may be referred to as VP2 of TTM-043. In some embodiments, an AAV particle of the present disclosure comprises a VP2 comprising the amino acid sequence of SEQ ID NO: 46. In some embodiments, an AAV particle of the present disclosure comprises a VP2 consisting of the amino acid sequence of SEQ ID NO: 46.

[0209] In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 45 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 thereto, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 96% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsidAttorney Docket No. 14640.0304-00304variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 98% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 45, wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 13 present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 449-466 of the amino acid sequence of SEQ ID NO: 45) and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45 (e.g., present at positions 590-596 of the amino acid sequence of SEQ ID NO: 45). In some embodiments, the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 45. In some embodiments, the amino acid sequence of SEQ ID NO: 45 may be referred to as VP1 of TTM-043. In some embodiments, an AAV particle of the present disclosure comprises a VP1 comprising the amino acid sequence of SEQ ID NO: 45. In some embodiments, anAttorney Docket No. 14640.0304-00304AAV particle of the present disclosure comprises a VP1 consisting of the amino acid sequence of SEQ ID NO: 45.

[0210] In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant that is encoded by the nucleotide sequence of SEQ ID NO: 48 or a nucleotide sequence that is 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%) identical thereto. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 80% identical to the nucleotide sequence of SEQ ID NO: 48. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 90% identical to the nucleotide sequence of SEQ ID NO: 48. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 95% identical to the nucleotide sequence of SEQ ID NO: 48. In some embodiments, the AAV capsid variant is encoded by a nucleotide sequence that is at least 99% identical to the nucleotide sequence of SEQ ID NO: 48. In some embodiments, the AAV capsid variant is encoded by the nucleotide sequence of SEQ ID NO: 48.

[0211] In some embodiments, an AAV particle of the present disclosure comprises an amino acid sequence provided in Table 10. In some embodiments, an AAV particle of the present disclosure comprises amino acids 2-742 of an amino acid sequence provided in Table 10. In some embodiments, an AAV particle of the present disclosure comprises an amino acid sequence encoded by a nucleotide sequence of Table 11. In some embodiments, an AAV particle of the present disclosure is encoded by a nucleotide sequence of Table 11.

[0212] In some embodiments, the AAV particle comprises the AAV capsid variant TTM-027.

[0213] In some embodiments, the AAV particle comprises the AAV capsid variant TTM-003.

[0214] In some embodiments, the AAV particle comprises the AAV capsid variant TTM-043.Table 10. Exemplary Capsid Amino Acid SequencesName SEQ Amino Acid SequenceID NO.TTM-003 4 MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYK VP1 YLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQE RLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEP DSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHAttorney Docket No. 14640.0304-00304Name SEQ Amino Acid SequenceID NO.LYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNW GFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLG SAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTG NNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTERVSGSPHS KAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWP GASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLI FGKQGTGRDNVDA DKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMV WQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPAD PPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKS NN VE FAVNT E GVY S E P RP I GT RYLT RNL TTM-003 26 TAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPI VP2 GEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRV ITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTST VQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQY LYYLSKTERVSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQR VSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSG SLIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQ AQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMK HPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENS KRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL TTM-003 27 MASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYN VP3 NHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINN NWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYV LGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLR TGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTERVSGSP HSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFA WPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLI FGKQGTGRDNV DADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPG MVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVP ADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYY KSNNVEFAVNTEGVYSEPRPIGTRYLTRNL TTM-027 1 MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYK VP1 YLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQE RLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEP DSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMA SGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNH LYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNW GFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLG SAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTG NNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTENVSGSPHS KAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWP GASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLI FGKQGTGRDNVDA DKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMV WQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPAD PPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKS NN VE FAVNT E GVY S E P RP I GT RYLT RNL TTM-027 24 TAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPI VP2 GEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTAttorney Docket No. 14640.0304-00304Name SEQ Amino Acid SequenceID NO.VQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQY LYYLSKTENVSGSPHSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQR VSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSG SLIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQ AQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMK HPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENS KRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL TTM-027 25 MASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYN VP3 NHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINN NWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYV LGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLR TGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTENVSGSP HSKAQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFA WPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLI FGKQGTGRDNV DADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPG MVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVP ADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYY KSNNVEFAVNTEGVYSEPRPIGTRYLTRNL TTM-043 45 MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYK VP1 YLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQE RLKEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEP DSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMA SGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNH LYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNW GFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLG SAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTG NNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGHDSPHK SGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWP GASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLI FGKQGTGRDNVDA DKVMITNEEEIKTTNPVATESYGQVATNRQTALQLAQTGWVQNQGILPGMV WQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPAD PPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKS NN VE FAVNT E GVY S E P RP I GT RYLT RNL TTM-043 46 TAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPI VP2 GEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRV ITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFH CHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTST VQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRS SFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQY LYYLSKTINGHDSPHKSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQR VSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSG SLIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNRQTALQL AQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMK HPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENS KRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL TTM-043 47 MASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYN VP3 NHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINN NWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYV LGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLR TGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGHDSPHKSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAAttorney Docket No. 14640.0304-00304Name SEQ Amino Acid SequenceID NO.WPGASSWALNGRNSLMNPGPAMASHKEGEDRFFPLSGSLI FGKQGTGRDNV DADKVMITNEEEIKTTNPVATESYGQVATNRQTALQLAQTGWVQNQGILPG MVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVP ADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNLTable 11. Exemplary Capsid Nucleotide SequencesName SEQ Nucleotide SequenceID NO.TTM-003 3 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACAACCTTAGTGAAGGAAT TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC AAAGCCTGGACCGACTAATGAATCCACTCATCGATCAGTATCTGTATTACTTGAGT AAGACGGAGCGTGTGTCTGGTTCTCCGCATTCTAAGGCGCAGAATCAGCAGACGTT GAAGTTTTCGGTAGCTGGTCCTAGCAACATGGCTGTCCAGGGAAGAAACTACATAC CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC AACGGAGTCCTATGGACAAGTGGCCACAAACCACCAGAGTGCCCAAGCACAGGCGC AGACCGGCTGGGTTCAAAACCAAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT GAACCCCGCCCCATTGGCACGCGGTATTTAACGAGGAACTTA TTM-027 2 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACAACCTTAGTGAAGGAAT TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGCAACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCAAttorney Docket No. 14640.0304-00304Name SEQ Nucleotide SequenceID NO.CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGA TTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCG TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC AAAGCCTGGACCGACTAATGAATCCACTCATCGACCAATACTTGTACTATCTCTCT AAGACT GAGAAT GT GAGCGGGAGCCCT CATAGCAAGGCT CAGAAT CAGCAGACT CT AAAATTCAGTGTGGCCGGACCCAGCAACATGGCTGTCCAGGGAAGAAACTACATAC CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT GATGAATCCTGGACCTGCTATGGCCAGCCACAAAGAAGGAGAGGACCGTTTCTTTC CTTTGTCTGGATCTTTAATTTTTGGCAAACAAGGAACTGGAAGAGACAACGTGGAT GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC AACGGAGTCCTATGGACAAGTGGCCACAAACCACCAGAGTGCCCAAGCACAGGCGC AGACCGGCTGGGTTCAAAACCAAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGT GAACCCCGCCCCATTGGCACCAGATACCTGACTCGTAATCTG TTM-043 48 ATGGCTGCCGATGGTTATCTTCCAGATTGGCTCGAGGACAACCTTAGTGAAGGAAT TCGCGAGTGGTGGGCTTTGAAACCTGGAGCCCCTCAACCCAAGGCAAATCAACAAC ATCAAGACAACGCTCGAGGTCTTGTGCTTCCGGGTTACAAATACCTTGGACCCGGC AACGGACTCGACAAGGGGGAGCCGGTCAACGCAGCAGACGCGGCGGCCCTCGAGCA CGACAAGGCCTACGACCAGCAGCTCAAGGCCGGAGACAACCCGTACCTCAAGTACA ACCACGCCGACGCCGAGTTCCAGGAGCGGCTCAAAGAAGATACGTCTTTTGGGGGC AACCTCGGGCGAGCAGTCTTCCAGGCCAAAAAGAGGCTTCTTGAACCTCTTGGTCT GGTTGAGGAAGCGGCTAAGACGGCTCCTGGAAAGAAGAGGCCTGTAGAGCAGTCTC CTCAGGAACCGGACTCCTCCGCGGGTATTGGCAAATCGGGTGCACAGCCCGCTAAA AAGAGACTCAATTTCGGTCAGACTGGCGACACAGAGTCAGTCCCAGACCCTCAACC AATCGGAGAACCTCCCGCAGCCCCCTCAGGTGTGGGATCTCTTACAATGGCTTCAG GTGGTGGCGCACCAGTGGCAGACAATAACGAAGGTGCCGATGGAGTGGGTAGTTCC TCGGGAAATTGGCATTGCGATTCCCAATGGCTGGGGGACAGAGTCATCACCACCAG CACCCGAACCTGGGCCCTGCCCACCTACAACAATCACCTCTACAAGCAAATCTCCA ACAGCACATCTGGAGGATCTTCAAATGACAACGCCTACTTCGGCTACAGCACCCCC TGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTCTCACCACGTGACTGGCA GCGACTCATCAACAACAACTGGGGATTCCGGCCTAAGCGACTCAACTTCAAGCTCT TCAACATTCAGGTCAAAGAGGTTACGGACAACAATGGAGTCAAGACCATCGCCAAT AACCTTACCAGCACGGTCCAGGTCTTCACGGACTCAGACTATCAGCTCCCGTACGT GCTCGGGTCGGCTCACGAGGGCTGCCTCCCGCCGTTCCCAGCGGACGTTTTCATGATTCCTCAGTACGGGTATCTGACGCTTAATGATGGAAGCCAGGCCGTGGGTCGTTCGAttorney Docket No. 14640.0304-00304Name SEQ Nucleotide SequenceID NO.TCCTTTTACTGCCTGGAATATTTCCCGTCGCAAATGCTAAGAACGGGTAACAACTT CCAGTTCAGCTACGAGTTTGAGAACGTACCTTTCCATAGCAGCTACGCTCACAGCC AAAGCCTGGACCGACTAATGAATCCACTCATCGACCAATACTTGTATTACTTGAGT AAAACAATTAACGGACACGACAGCCCACACAAAAGCGGACAAAACCAACAGACCTT GAAGTTTTCGGTAGCTGGTCCTAGCAACATGGCTGTCCAGGGAAGAAACTACATAC CTGGACCCAGCTACCGACAACAACGTGTCTCAACCACTGTGACTCAAAACAACAAC AGCGAATTTGCTTGGCCTGGAGCTTCTTCTTGGGCTCTCAATGGACGTAATAGCTT GATGAATCCTGGACCTGCTATGGCGTCTCATAAAGAAGGGGAGGATCGTTTCTTTC CTTTGTCTGGATCTTTAATTTTTGGTAAGCAGGGTACAGGTAGAGACAACGTGGAT GCGGACAAAGTCATGATAACCAACGAAGAAGAAATTAAAACTACTAACCCGGTAGC AACGGAGTCCTATGGACAAGTGGCTACCAACAGGCAGACCGCCCTGCAGCTGGCGC AGACCGGCTGGGTTCAAAACCAAGGAATACTTCCGGGTATGGTTTGGCAGGACAGA GATGTGTACCTGCAAGGACCCATTTGGGCCAAAATTCCTCACACGGACGGCAACTT TCACCCTTCTCCGCTGATGGGAGGGTTTGGAATGAAGCACCCGCCTCCTCAGATCC TCATCAAAAACACACCTGTACCTGCGGATCCTCCAACGGCCTTCAACAAGGACAAG CTGAACTCTTTCATCACCCAGTATTCTACTGGCCAAGTCAGCGTGGAGATCGAGTG GGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCGGAGATCCAGTACACTTCCA ACTATTACAAGTCTAATAATGTTGAATTTGCTGTTAATACTGAAGGTGTATATAGTGAACCCCGCCCCATTGGCACCCGGTATTTAACGAGGAACTTA

[0215] It is common for a first amino acid (AA1), e.g., a first methionine (Metl), encoded by a capsid gene to be expressed in capsid proteins but cleaved during or after polypeptide synthesis by protein processing enzymes such as Met-aminopeptidases. This “Metl / AAl-clipping” process often correlates with a corresponding acetylation of the second amino acid in the polypeptide sequence (e.g., alanine, valine, serine, threonine, etc.). Metl / AAl -clipping commonly occurs with VP1 and VP3 but can also occur with VP2.

[0216] Where the Metl / AAl -clipping is incomplete, a mixture of VP capsid proteins comprising the viral capsid may be produced, some of which include a Metl / AAl amino acid (Met+ / AA+) and some of which may lack a Metl / AAl amino acid as a result of Metl / AAl-clipping (Metl- / AA1-). For further discussion regarding Metl / AAl -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 relevant contents of each of which are incorporated herein by reference in their entirety.

[0217] According to the present disclosure, references to capsid proteins, e.g., of AAV capsid variants, is not limited to either clipped (Metl- / AA1-) or unclipped (Metl+ / AA1+) and may refer to a mixture of clipped and unclipped capsid proteins. A direct reference to a capsid protein whether by VP1, VP2, or VP3 designation or by SEQ ID NO may encompassAttorney Docket No. 14640.0304-00304VP capsid proteins that include Metl / AAl (Metl+ / AA1+) as well as corresponding VP capsid proteins without Metl / AAl as a result of clipping (Metl- / AA1-).

[0218] Accordingly, in some embodiments, an AAV particle of the present disclosure comprises a Metl / AAl -clipped VP1, VP2, and / or VP3 of an AAV9 capsid variant described herein. In some embodiments, an AAV particle of the present disclosure comprises a Metl / AAl-clipped VP1, VP2, and / or VP3 of TTM-027. In some embodiments, an AAV particle of the present disclosure comprises a Metl / AAl-clipped VP1, VP2, and / or VP3 of TTM-003. In some embodiments, an AAV particle of the present disclosure comprises a Metl / AAl-clipped VP1, VP2, and / or VP3 ofTTM-043.

[0219] In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-742 of the amino acid sequence of SEQ ID NO: 1, wherein amino acid 2 of SEQ ID NO: 1 is optionally acetylated. In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-605 of the amino acid sequence of SEQ ID NO: 24, wherein amino acid 2 of SEQ ID NO: 24 is optionally acetylated. In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-540 of the amino acid sequence of SEQ ID NO: 25, wherein amino acid 2 of SEQ ID NO: 25 is optionally acetylated.

[0220] In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-742 of the amino acid sequence of SEQ ID NO: 4, wherein amino acid 2 of SEQ ID NO: 4 is optionally acetylated. In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-605 of the amino acid sequence of SEQ ID NO: 26, wherein amino acid 2 of SEQ ID NO: 26 is optionally acetylated. In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-540 of the amino acid sequence of SEQ ID NO: 27, wherein amino acid 2 of SEQ ID NO: 27 is optionally acetylated.

[0221] In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-742 of the amino acid sequence of SEQ ID NO: 45, wherein amino acid 2 of SEQ ID NO: 45 is optionally acetylated. In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-605 of the amino acid sequence of SEQ ID NO: 46, wherein amino acid 2 of SEQ ID NO: 46 is optionally acetylated. In some embodiments, an AAV particle of the present disclosure comprises an AAV capsid variant comprising amino acids 2-540 of the amino acidAttorney Docket No. 14640.0304-00304sequence of SEQ ID NO: 47, wherein amino acid 2 of SEQ ID NO: 47 is optionally acetylated.C. Exemplary AAV Particles

[0222] In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 15 and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising, in 5’ to 3’ order, the nucleotide sequence of SEQ ID NO: 18, the nucleotide sequence of SEQ ID NO: 15, and the nucleotide sequence of SEQ ID NO: 19.

[0223] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 16 and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising, in 5’ to 3’ order, the nucleotide sequence of SEQ ID NO: 18, the nucleotide sequence of SEQ ID NO: 16 and the nucleotide sequence of SEQ ID NO: 19.

[0224] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising, in 5’ to 3’ order, a 5’ ITR comprising the nucleotide sequence of SEQ ID NO: 17; a promoter comprising the nucleotide sequence of SEQ ID NO: 18; a CDKL5 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; a WPRE comprising the nucleotide sequence of SEQ ID NO: 19; optionally a nucleotide sequence encoding a microRNA binding site, comprising the nucleotide sequence of SEQ ID NO: 6; a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21. In some embodiments, the nucleotide sequence encoding the microRNA binding site comprises the nucleotide sequence of SEQ ID NO: 7.Attorney Docket No. 14640.0304-00304

[0225] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant is TTM-027) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 30.

[0226] In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-027 and the viral genome (e.g., recombinant viral genome) of Construct 1. In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-027 and the viral genome (e.g., recombinant viral genome) of Construct 2. In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-027 and the viral genome (e.g., recombinant viral genome) of Construct 3.

[0227] In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the AAV capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 15 and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the AAV capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising, in 5’ to 3’ order, the nucleotide sequence of SEQ ID NO: 18, the nucleotide sequence of SEQ ID NO: 15, and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the AAV capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 16 and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the AAV capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising, in 5’ to 3’ order, the nucleotide sequence of SEQ ID NO: 18, the nucleotide sequence of SEQ ID NO: 16, and the nucleotide sequence of SEQ ID NO: 19.Attorney Docket No. 14640.0304-00304

[0228] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising a 5’ ITR comprising the nucleotide sequence of SEQ ID NO: 17; a promoter comprising the nucleotide sequence of SEQ ID NO: 18; a CDKL5 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; a WPRE comprising the nucleotide sequence of SEQ ID NO: 19; optionally a nucleotide sequence encoding a microRNA binding site, comprising the nucleotide sequence of SEQ ID NO: 6; a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21. In some embodiments, the nucleotide sequence encoding the microRNA binding site comprises the nucleotide sequence of SEQ ID NO: 7.

[0229] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., the capsid variant is TTM-003) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 30.

[0230] In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-003 and the viral genome (e.g., recombinant viral genome) of Construct 1. In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-003 and the viral genome (e.g., recombinant viral genome) of Construct 2. In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-003 and the viral genome (e.g., recombinant viral genome) of Construct 3.

[0231] In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the AAV capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 15 and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the AAV capsid variant is TTM-043) and a viral genome (e.g.,Attorney Docket No. 14640.0304-00304recombinant viral genome) comprising, in 5’ to 3’ order, the nucleotide sequence of SEQ ID NO: 18, the nucleotide sequence of SEQ ID NO: 15, and the nucleotide sequence of SEQ ID NO: 19.

[0232] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the AAV capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 16 and the nucleotide sequence of SEQ ID NO: 19. In some embodiments, the present disclosure provides an AAV particle that comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the AAV capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising, in 5’ to 3’ order, the nucleotide sequence of SEQ ID NO: 18, the nucleotide sequence of SEQ ID NO: 16, and the nucleotide sequence of SEQ ID NO: 19.

[0233] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising a 5’ ITR comprising the nucleotide sequence of SEQ ID NO: 17; a promoter comprising the nucleotide sequence of SEQ ID NO: 18; a CDKL5 -encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16; a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19; optionally a nucleotide sequence encoding a microRNA binding site, comprising the nucleotide sequence of SEQ ID NO: 6; a polyA sequence comprising the nucleotide sequence of SEQ ID NO: 20; and a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21. In some embodiments, the nucleotide sequence encoding the microRNA binding site comprises the nucleotide sequence of SEQ ID NO: 7.

[0234] In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 22. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 23. In some embodiments, the AAV particle comprises an AAV capsid variant comprising the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the capsid variant is TTM-043) and a viral genome (e.g., recombinant viral genome) comprising the nucleotide sequence of SEQ ID NO: 30.Attorney Docket No. 14640.0304-00304

[0235] In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-043 and the viral genome (e.g., recombinant viral genome) of Construct 1. In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-043 and the viral genome (e.g., recombinant viral genome) of Construct 2. In some embodiments, the AAV particle comprises the AAV capsid variant of TTM-043 and the viral genome (e.g., recombinant viral genome) of Construct 3.D. Tropism and Biodistribution Properties

[0236] 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. In some embodiments, the present disclosure provides AAV particles that demonstrate enhanced tropism for a target cell type, tissue or organ, wherein the AAV particles comprise the amino acid sequence of SEQ ID NO: 1, 24, or 25 (e.g., the AAV capsid variant TTM-027), the amino acid sequence of SEQ ID NO: 4, 26, or 27 (e.g., AAV capsid variant TTM-003), or the amino acid sequence of SEQ ID NO: 45, 46, or 47 (e.g., the AAV capsid variant TTM-043). As a non-limiting example, an AAV particle of the disclosure may have enhanced tropism for cells and tissues o...

Claims

Attorney Docket No. 14640.0304-00304ClaimsWe claim:

1. An adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE);wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25 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 thereto; andwherein the AAV9 capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 5) present at amino acids corresponding to positions 254-259 of the amino acid sequence of SEQ ID NO: 25, the amino acid E at position 249 of the amino acid sequence of SEQ ID NO: 25, and the amino acid V at position 251 of the amino acid sequence of SEQ ID NO: 25.

2. An adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE);wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 25 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 thereto; andwherein the AAV9 capsid variant comprises the amino acid sequence of ENVSGSPHSKA (SEQ ID NO: 8) present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 25, optionally wherein the AAV9 capsid variant comprises the amino acid sequence of KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 25.

3. The AAV particle of claim 1 or claim 2, wherein the AAV9 capsid variant comprises:(i) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 1, wherein theAttorney Docket No. 14640.0304-00304amino acid sequence comprises ENVSGSPHSKA (SEQ ID NO: 8) present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 1, optionally wherein the amino acid sequence comprises KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 1; and / or(ii) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 24, wherein the amino acid sequence comprises ENVSGSPHSKA (SEQ ID NO: 8) present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 24, optionally wherein the amino acid sequence comprises KTENVSGSPHSKAQNQQT (SEQ ID NO: 9) present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 24.

4. The AAV particle of any one of claims 1-3, wherein the AAV9 capsid variant comprises:(i) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 1;(ii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 24; and / or(iii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 25.

5. The AAV particle of any one of claims 1-4, wherein the AAV9 capsid variant comprises:(i) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 1;(ii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 24; and / or(iii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 25.Attorney Docket No. 14640.0304-003046. The AAV particle of any one of claims 1-5, wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 1;(ii) the amino acid sequence of SEQ ID NO: 24; and / or(iii) the amino acid sequence of SEQ ID NO: 25.

7. An adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE);wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27 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 thereto; andwherein the AAV9 capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 5) present at amino acids corresponding to positions 254-259 of the amino acid sequence of SEQ ID NO: 27, the amino acid E at position 249 of the amino acid sequence of SEQ ID NO: 27, the amino acid R at position 250 numbered according to the amino acid sequence of SEQ ID NO: 27, and the amino acid V at position 251 of the amino acid sequence of SEQ ID NO: 27.

8. An adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE);wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 27 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 thereto; andwherein the AAV9 capsid variant comprises the amino acid sequence of ERVSGSPHSKA (SEQ ID NO: 10) present at amino acids corresponding to positions 249-259 of the amino acid sequence of SEQ ID NO: 27, optionally wherein the AAV9 capsid variant comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 27.Attorney Docket No. 14640.0304-003049. The AAV particle of claim 7 or claim 8, wherein the AAV9 capsid variant comprises:(i) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 4, wherein the amino acid sequence comprises ERVSGSPHSKA (SEQ ID NO: 10) present at amino acids corresponding to positions 451-461 of the amino acid sequence of SEQ ID NO: 4, optionally wherein the amino acid sequence comprises KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 4; and / or(ii) an amino acid sequence that is 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 26, wherein the amino acid sequence comprises ERVSGSPHSKA (SEQ ID NO: 10) present at amino acids corresponding to positions 314-324 of the amino acid sequence of SEQ ID NO: 26, optionally wherein the amino acid sequence comprises KTERVSGSPHSKAQNQQT (SEQ ID NO: 11) present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 26.

10. The AAV particle of any one of claims 7-9, wherein the AAV9 capsid variant comprises:(i) an amino acid sequence that is at least 95% (at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 4;(ii) an amino acid sequence that is at least 95% (at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 26; and / or(iii) an amino acid sequence that is at least 95% (at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 27.

11. The AAV particle of any one of claims 7-10, wherein the AAV9 capsid variant comprises:(i) an amino acid sequence that is at least 99% identical to SEQ ID NO: 4;Attorney Docket No. 14640.0304-00304(ii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 26; and / or(iii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 27.

12. The AAV particle of any one of claims 7-11, wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 4;(ii) the amino acid sequence of SEQ ID NO: 26; and / or(iii) the amino acid sequence of SEQ ID NO: 27.

13. An adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE);wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47 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 thereto; andwherein the AAV9 capsid variant comprises HDSPHK (SEQ ID NO: 12) present at amino acids 252-257 of the amino acid sequence of SEQ ID NO: 47 and comprises the amino acid R at position 388, the amino acid T at position 390, the amino acid L at position 392, the amino acid Q at position 393, and the amino acid L at position 394, each numbered according to the amino acid sequence of SEQ ID NO: 47.

14. An adeno-associated virus (AAV) particle comprising an AAV9 capsid variant and a recombinant viral genome comprising a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE);wherein the AAV9 capsid variant comprises the amino acid sequence of SEQ ID NO: 47 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 thereto; andwherein the AAV9 capsid variant comprises HDSPHK (SEQ ID NO: 12) present at amino acids corresponding to positions 252-257 of the amino acid sequence of SEQ ID NO:Attorney Docket No. 14640.0304-0030447 and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 388-394 of the amino acid sequence of SEQ ID NO: 47; optionally wherein the AAV9 capsid variant comprises the amino acid sequence of KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) present at amino acids corresponding to positions 247-264 of the amino acid sequence of SEQ ID NO: 47.

15. The AAV particle of claim 13 or claim 14, wherein the AAV9 capsid variant comprises:(i) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 45, wherein the amino acid sequence comprises HDSPHK (SEQ ID NO: 12) present at amino acids corresponding to positions 454-459 of the amino acid sequence of SEQ ID NO: 45 and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 590-596 of the amino acid sequence of SEQ ID NO: 45, optionally wherein the amino acid sequence comprises KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) present at amino acids corresponding to positions 449-466 of the amino acid sequence of SEQ ID NO: 45; and / or(ii) 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%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 46, wherein the amino acid sequence comprises HDSPHK (SEQ ID NO: 12) present at amino acids corresponding to positions 317-322 of the amino acid sequence of SEQ ID NO: 46 and comprises the amino acid sequence of RQTALQL (SEQ ID NO: 49) present at amino acids corresponding to positions 453-459 of the amino acid sequence of SEQ ID NO: 46, optionally wherein the amino acid sequence comprises KTINGHDSPHKSGQNQQT (SEQ ID NO: 13) present at amino acids corresponding to positions 312-329 of the amino acid sequence of SEQ ID NO: 46.

16. The AAV particle of any one of claims 13-15, wherein the AAV9 capsid variant comprises:(i) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 45;Attorney Docket No. 14640.0304-00304(ii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 46; and / or(iii) an amino acid sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the amino acid sequence of SEQ ID NO: 47.

17. The AAV particle of any one of claims 13-16, wherein the AAV9 capsid variant comprises:(i) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 45;(ii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 46; and / or(iii) an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO: 47.

18. The AAV particle of any one of claims 13-17, wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 45;(ii) the amino acid sequence of SEQ ID NO: 46; and / or(iii) the amino acid sequence of SEQ ID NO: 47.

19. The AAV particle of any one of claims 1-18, wherein the recombinant viral genome encodes a wildtype CDKL5 protein.

20. The AAV particle of any one of claims 1-19, wherein the recombinant viral genome encodes a human CDKL5 protein.

21. The AAV particle of claim 19 or claim 20, wherein the encoded CDKL5 comprises the amino acid sequence of SEQ ID NO: 14 or any one of SEQ ID NOs: 34-37.

22. The AAV particle of any one of claims 1-21, wherein the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide sequence that is at least 70% (e.g., at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, atAttorney Docket No. 14640.0304-00304least 76%, at least 77%, at least 78%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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.

23. The AAV particle of any one of claims 1-22, wherein the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide 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 thereto.

24. The AAV particle of any one of claims 1-23, wherein the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15.

25. The AAV particle of any one of claims 1-21, wherein the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16 or a nucleotide sequence that is at least 98% (e.g., at least 98% or at least 99%) identical thereto.

26. The AAV particle of claim 25, wherein the CDKL5-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16.

27. The AAV particle of any one of claims 1-26, wherein the WPRE is positioned 3’ relative to the CDKL5 -encoding sequence.

28. The AAV particle of any one of claims 1-27, wherein the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or any one of SEQ ID NOs: 58-62, or a nucleotide 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 thereto.

29. The AAV particle of any one of claims 1-28, wherein the WPRE comprises the nucleotide sequence of SEQ ID NO: 19.

30. The AAV particle of any one of claims 1-29, wherein the recombinant viral genome further comprises a promoter operably linked to the CDKL5-encoding sequence.Attorney Docket No. 14640.0304-0030431. The AAV particle of claim 30, wherein the promoter is a human synapsin 1 (hSYNl) promoter or a chicken P-actin hybrid (CBh) promoter.

32. The AAV particle of claim 30 or claim 31, wherein the promoter is a hSYNl promoter.

33. The AAV particle of any one of claims 30-32, wherein the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto.

34. The AAV particle of any one of claims 30-33, wherein the promoter comprises the nucleotide sequence of SEQ ID NO: 18.

35. The AAV particle of any one of claims 1-34, wherein the recombinant viral genome further comprises a polyadenylation (poly A) sequence.

36. The AAV particle of claim 35, wherein the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto.

37. The AAV particle of claim 35 or claim 36, wherein the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20.

38. The AAV particle of any one of claims 1-37, wherein the recombinant viral genome further comprises at least one inverted terminal repeat (ITR).

39. The AAV particle of claim 38, wherein the at least one ITR comprises a 5’ ITR and a 3’ ITR.

640. The AAV particle of claim 39, wherein the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto.Attorney Docket No. 14640.0304-0030441. The AAV particle of claim 39 or claim 40, wherein the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17.

42. The AAV particle of any one of claims 39-41, wherein the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical thereto.

43. The AAV particle of any one of claims 39-42, wherein the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21.

44. The AAV particle of any one of claims 1-43, wherein the recombinant viral genome further comprises a nucleotide sequence encoding one or more microRNA (miR) binding sites, optionally wherein the one or more miR binding sites reduces or prevents expression of CDKL5 in dorsal root ganglia.

45. The AAV particle of claim 44, wherein the one or more miR binding sites comprises one, two, three, or four miR183 binding sites.

46. The AAV particle of claim 44 or claim 45, wherein the recombinant viral genome comprises a nucleotide sequence encoding four miR183 binding sites, optionally wherein the four miR183 binding sites are identical.

47. The AAV particle of claim 46, wherein each of the four miR183 binding sites is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 6.

48. The AAV particle of claim 47, wherein each of the four miR183 binding sites is encoded by the nucleotide sequence of SEQ ID NO: 6.

49. The AAV particle of any one of claims 46-48, wherein the miR183 binding sites are separated by a spacer, optionally wherein the spacer is encoded by the nucleotide sequence GATAGTTA.

50. The AAV particle of any one of claims 1-49, wherein the recombinant viral genome further comprises a nucleotide sequence encoding a microRNA183 (miR183) binding siteAttorney Docket No. 14640.0304-00304series, wherein the nucleotide sequence encoding the miR183 binding site series comprises the nucleotide sequence of SEQ ID NO: 7 or a nucleotide 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 thereto.

51. The AAV particle of claim 50, wherein the nucleotide sequence encoding the miRl 83 binding site series comprises the nucleotide sequence of SEQ ID NO: 7.

52. The AAV particle of claim 51, wherein the nucleotide sequence encoding the miRl 83 binding site series consists of the nucleotide sequence of SEQ ID NO: 7.

53. The AAV particle of any one of claims 1-52, wherein the recombinant viral genome comprises, in 5’ to 3’ order:a) a 5’ inverted terminal repeat (ITR);b) a promoter;c) the CDKL5 -encoding sequence;d) the WPRE;e) a polyadenylation (poly A) sequence; andf) a 3’ ITR.

54. The AAV particle of claim 53, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide 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 thereto;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto;Attorney Docket No. 14640.0304-00304e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; and / orf) the 3’ ITR comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

55. The AAV particle of claim 53 or claim 54, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or a nucleotide 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 thereto;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; andf) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

56. The AAV particle of any one of claims 53-55, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto;Attorney Docket No. 14640.0304-00304c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19 or a nucleotide 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 thereto;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; andf) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

57. The AAV particle of any one of claims 53-56, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18 or a nucleotide 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 thereto;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; andf) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

58. The AAV particle of any one of claims 53-57, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17 or a nucleotide 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 thereto;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18;Attorney Docket No. 14640.0304-00304c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; andf) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

59. The AAV particle of any one of claims 53-58, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; andf) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21 or a nucleotide 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 thereto.

60. The AAV particle of any one of claims 53-59, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20 or a nucleotide 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 thereto; andf) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21.Attorney Docket No. 14640.0304-0030461. The AAV particle of any one of claims 53-60, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 15;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20; and f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21.

62. The AAV particle of any one of claims 53-61, wherein:a) the 5’ ITR comprises the nucleotide sequence of SEQ ID NO: 17;b) the promoter comprises the nucleotide sequence of SEQ ID NO: 18;c) the CDKL5 -encoding sequence comprises the nucleotide sequence of SEQ ID NO: 16;d) the WPRE comprises the nucleotide sequence of SEQ ID NO: 19;e) the polyA sequence comprises the nucleotide sequence of SEQ ID NO: 20; and f) the 3’ ITR sequence comprises the nucleotide sequence of SEQ ID NO: 21.

63. The AAV particle of any one of claims 1-62, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22 or a nucleotide 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 thereto.

64. The AAV particle of claim 63, wherein the recombinant viral genome comprises a nucleotide sequence that is at least 95% (e.g., at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to SEQ ID NO: 22.

65. The AAV particle of claim 63 or claim 64, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22.

66. The AAV particle of claim 63 or claim 64, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23.Attorney Docket No. 14640.0304-0030467. The AAV particle of any one of claims 1-66, wherein the recombinant viral genome further comprises a nucleotide sequence encoding one or more microRNA (miR) binding sites, optionally wherein the one or more miR binding sites reduces or prevents expression of CDKL5 in dorsal root ganglia.

68. The AAV particle of claim 67, wherein the nucleotide sequence encoding the one or more miR binding sites comprises the nucleotide sequence of SEQ ID NO: 6.

69. The AAV particle of claim 67 or claim 68, wherein the nucleotide sequence encoding the one or more miR binding sites encodes four miR binding sites, wherein each of the four miR binding sites is encoded by a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 6.

70. The AAV particle of any one of claims 1-69, wherein the recombinant viral genome further comprises a nucleotide sequence encoding a microRNA (miR) binding site series, wherein the nucleotide sequence encoding the miR binding site series comprises the nucleotide sequence of SEQ ID NO: 7.

71. The AAV particle of any one of claims 53-61, 63, 64, or 67-70, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

72. An adeno-associated virus (AAV) particle comprising a recombinant viral genome and an AAV9 capsid variant,wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 1;(ii) the amino acid sequence of SEQ ID NO: 24; and / or(iii) the amino acid sequence of SEQ ID NO: 25; andwherein the recombinant viral genome comprises:a) a 5’ inverted terminal repeat (ITR) comprising the nucleotide sequence of SEQ ID NO: 17;b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18;c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;Attorney Docket No. 14640.0304-00304d) a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19;e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6;f) a polyadenylation (poly A) sequence comprising the nucleotide sequence of SEQ ID NO: 20; andg) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

73. The AAV particle of claim 69, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22.

74. The AAV particle of claim 69, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23.

75. The AAV particle of claim 69, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

76. An adeno-associated virus (AAV) particle comprising a recombinant viral genome and an AAV9 capsid variant,wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 4;(ii) the amino acid sequence of SEQ ID NO: 26; and / or(iii) the amino acid sequence of SEQ ID NO: 27; andwherein the recombinant viral genome comprises:a) a 5’ inverted terminal repeat (5’ ITR) comprising the nucleotide sequence of SEQ ID NO: 17;b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18;c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19;Attorney Docket No. 14640.0304-00304e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6;f) a polyadenylation (poly A) sequence comprising the nucleotide sequence of SEQ ID NO: 20; andg) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

77. The AAV particle of claim 73, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22.

78. The AAV particle of claim 73, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23.

79. The AAV particle of claim 73, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

80. An adeno-associated virus (AAV) particle comprising a recombinant viral genome and an AAV9 capsid variant,wherein the AAV9 capsid variant comprises:(i) the amino acid sequence of SEQ ID NO: 45;(ii) the amino acid sequence of SEQ ID NO: 46; and / or(iii) the amino acid sequence of SEQ ID NO: 47; andwherein the recombinant viral genome comprises:a) a 5’ inverted terminal repeat (5’ ITR) comprising the nucleotide sequence of SEQ ID NO: 17;b) a promoter comprising the nucleotide sequence of SEQ ID NO: 18;c) a cyclin-dependent kinase-like 5 (CDKL5)-encoding sequence comprising the nucleotide sequence of SEQ ID NO: 15 or SEQ ID NO: 16;d) a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) comprising the nucleotide sequence of SEQ ID NO: 19;e) optionally a nucleotide sequence encoding a microRNA (miR) binding site, wherein the nucleotide sequence encoding the miR binding site comprises the nucleotide sequence of SEQ ID NO: 6;Attorney Docket No. 14640.0304-00304f) a polyadenylation (poly A) sequence comprising the nucleotide sequence of SEQ ID NO: 20; andg) a 3’ ITR sequence comprising the nucleotide sequence of SEQ ID NO: 21.

81. The AAV particle of claim 80, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22.

82. The AAV particle of claim 80, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23.

83. The AAV particle of claim 80, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.

84. A cell comprising the AAV particle of any one of claims 1-83.

85. The cell of claim 84, wherein the cell is a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

86. A method of making the AAV particle of any one of claims 1-83, the method comprising:(i) providing a cell comprising the recombinant viral genome comprising a CDKL5-encoding sequence and a nucleic acid encoding the AAV9 capsid variant; and(ii) incubating the cell under conditions suitable to encapsulate the recombinant viral genome in the AAV9 capsid variant;thereby making the AAV particle.

87. The method of claim 86, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 22.

88. The method of claim 86, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 23.

89. The method of claim 86, wherein the recombinant viral genome comprises the nucleotide sequence of SEQ ID NO: 30.Attorney Docket No. 14640.0304-0030490. The method of any one of claims 86-89, further comprising, prior to step (i), introducing a nucleic acid comprising the recombinant viral genome into the cell.

91. The method of any one of claims 86-90, further comprising, prior to step (i), introducing the nucleic acid encoding the AAV9 capsid variant into the cell.

92. The method of any one of claims 86-91, wherein the cell comprises a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

93. A pharmaceutical composition comprising the AAV particle of any one of claims 1-83 and a pharmaceutically acceptable excipient.

94. A method of delivering an AAV particle encoding a cyclin-dependent kinase-like 5 (CDKL5) to a cell, comprising administering an effective amount of the AAV particle of any one of claims 1-83 or the pharmaceutical composition of claim 93.

95. The method of claim 94, wherein the cell is in a subject.

96. The method of claim 95, wherein the subject has, has been diagnosed with having, or is at risk of having a CDKL5-related disorder.

97. A method of treating a subject having or diagnosed with having a CDKL5-related disorder, or at least one symptom thereof, comprising administering to the subject an effective amount of the AAV particle of any one of claims 1-83 or the pharmaceutical composition of claim 93.

98. The method of claim 96 or claim 97, wherein the CDKL5-related disorder is a CDKL5-related neurodegenerative or neuromuscular disorder.

99. The method of claim 98, wherein the CDKL5-related neurodegenerative or neuromuscular disorder is CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome.Attorney Docket No. 14640.0304-00304100. A method of treating a subject having or having or diagnosed with having a CDKL5-related disorder, or treating at least one symptom thereof, wherein the CDKL5-related disorder is CDKL5 deficiency disorder (CDD), comprising administering to the subject an effective amount of the AAV particle of any one of claims 1-83 or the pharmaceutical composition of claim 93.

101. The method of any one of claims 95-100, wherein the subject has one or more mutations in a CDKL5 gene.

102. The method of any one of claims 95-101, wherein the subject has a reduced level of CDKL5 activity as compared to a reference level in an individual who does not have a CDKL5-related disorder.

103. The method of any one of claims 97-102, wherein the treating results in prevention or progression of the disorder or at least one symptom thereof in the subject.

104. The method of any one of claims 97-103, wherein the treating results in amelioration of at least one symptom of the disorder in the subject, e.g., as indicated by one or more biomarkers.

105. The method of claim 104, wherein the one or more biomarkers comprises neurofilament light chain or a marker of CDKL5 activity, e.g., as measured by phosphorylation levels of substrate proteins, e.g., MECP2, or as measured by mass spectrometry.

106. The method of any one of claims 97-105, wherein the at least one symptom comprises epilepsy (e.g., early-onset epilepsy), autism, deficits in cognition, limited motor skills, sleep difficulties, visual impairment, low muscle tone, gastrointestinal reflux, behavioral symptoms (including episodes of laughing or crying that occur for what appears to be no reason, hypersensitivity to touch, and disrupted sleep), facial appearance changes (including microcephaly; a high, broad forehead; large, deep-set eyes; smaller-than-normal space between the nose and upper lip; an upturned nose; full lips and / or widely-spaced teeth), difficulties standing and walking, small, cold feet, lack of or poor eye contact, frequent sideways glances, cortical visual impairment or cortical blindness, bruxism, limited or absentAttorney Docket No. 14640.0304-00304speech, difficulties eating, stereotypies, limited ability to make small and / or focused hand movements, gastroesophageal reflux, constipation, or a combination thereof.

107. The method of any one of claims 95-106, wherein the subject is a human.

108. The method of any one of claims 95-107, wherein the AAV particle or the pharmaceutical composition is delivered to a cell, tissue, or region of the central nervous system (CNS) of the subject.

109. The method of claim 108, wherein the cell, tissue, or region of the CNS comprises: the parenchyma, cortex, substantia nigra, caudate cerebellum, striatum, corpus callosum, cerebellum, brain stem, caudate-putamen, thalamus, hippocampus, superior colliculus, spinal cord, or a combination thereof and / orneurons (e.g. glutamatergic neurons, GABAergic neurons, or a combination thereof).

110. The method of claim 108 or claim 109, wherein the cell, tissue, or region of the CNS is a cell, tissue, or region of the cortex (e.g., motor cortex), hippocampus, striatum, thalamus, or cerebellum, and / or wherein the cell of the CNS is a glutamatergic neuron and / or GABAergic neuron.

111. The method of any one of claims 95-110, wherein the AAV particle or pharmaceutical composition is delivered to the subject via intravenous administration.

112. The method of any one of claims 95-111, further comprising evaluating, e.g., measuring, the level of CDKL5 gene expression, CDKL5 mRNA expression, and / or CDKL5 protein expression in the subject, e.g., in a cell, tissue, or fluid of the subject.

113. The method of claim 112, wherein the level of CDKL5 protein expression is measured by an enzyme-linked immunosorbent assay (ELISA), a Western blot, or an immunohistochemistry assay.

114. The method of claim 112 or claim 113, wherein the evaluating the level of CDKL5 gene, mRNA, and / or protein expression is performed before and after administering the AAV particle or pharmaceutical composition, optionally wherein the subject’s level of CDKL5Attorney Docket No. 14640.0304-00304gene, mRNA, and / or protein expression before administration is compared to the subject’s level of CDKL5 gene, mRNA, and / or protein expression after administration.

115. The method of any one of claims 112-114, comprising evaluating the level of CDKL5 gene, mRNA, and / or protein expression in a cell or tissue of the CNS in the subject.

116. The method of claim 115, wherein the cell or tissue of the CNS comprises:the parenchyma, cortex, substantia nigra, caudate cerebellum, striatum, corpus callosum, cerebellum, brain stem, caudate-putamen, thalamus, hippocampus, superior colliculus, spinal cord, or a combination thereof and / orneurons (e.g. glutamatergic neurons, GABAergic neurons, or a combination thereof).

117. The method of claim 115 or claim 116, wherein the cell or tissue of the CNS is a cell or tissue of the cortex (e.g., motor cortex), hippocampus, striatum, thalamus, or cerebellum, and / or wherein the cell of the CNS is a glutamatergic neuron and / or GABAergic neuron.

118. The method of any one of claims 112-117, wherein the subject’s level of CDKL5 protein expression after administration of the AAV particle or pharmaceutical composition is increased relative to the subject’s level of CDKL5 protein expression before administration of the AAV particle or pharmaceutical composition.

119. The method of any one of claims 95-118, further comprising evaluating, e.g., measuring, the level of CDKL5 protein activity in the subject, e.g., in a cell or tissue of the subject.

120. The method of any one of claims 95-119, wherein administering the AAV particle or pharmaceutical composition to the subject results in an increase in:(i) the level of CDKL5 activity in a cell, tissue, or fluid (e.g., a cell or tissue of the CNS, e.g., the cortex, hippocampus, striatum, thalamus, cerebellum, glutamatergic neurons, GABAergic neurons, or a combination thereof) of the subject, relative to baseline and / or relative to the level of CDKL5 activity in a cell, tissue, or fluid of an individual with a CDKL5-related disorder who has not been administered the AAV particle or pharmaceutical composition;Attorney Docket No. 14640.0304-00304(ii) the number and / or level of recombinant viral genomes (VG) per cell level in a cell or tissue of the CNS (e.g., the cortex, hippocampus, striatum, thalamus, cerebellum, glutamatergic neurons, GABAergic neurons, or a combination thereof) of the subject, relative to the number and / or level of VG per cell in a peripheral cell or tissue of the subject; and / or (iii) the level of CDKL5 protein expression, CDKL5 mRNA expression, or CDKL5 gene expression in a cell or tissue (e.g., a cell or tissue of the CNS, e.g., the cortex, hippocampus, striatum, thalamus, cerebellum, glutamatergic neurons, GABAergic neurons, or a combination thereof) of the subject relative to baseline and / or relative to the level of CDKL5 protein expression, CDKL5 mRNA expression, or CDKL5 gene expression in a cell or tissue of an individual with a CDKL5-related disorder who has not been administered the AAV particle or pharmaceutical composition..

121. The method of any one of claims 97-120, further comprising administering to the subject at least one additional agent and / or therapy suitable for treating the CDKL5-related disorder or at least one symptom thereof.

122. The method of claim 121, wherein the at least one additional agent and / or therapy comprises one or more anti-epileptic drugs (e.g., bromide, clobazam, felbamate, ganaxolone, lamotrigine, levetiracetam, phenobarbital, topiramate, valproate, or a combination thereof).

123. The method of any one of claims 95-122, further comprising administering an immunosuppressant to the subject.

124. The method of claim 123, wherein the immunosuppressant comprises adrenocorticotropic hormone, a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), eculizumab hydroxychloroquine, mycophenolate mofetil, rapamycin, rituximab, and / or tacrolimus.

125. The AAV particle of any one of claims 1-83 or the pharmaceutical composition of claim 93, for use in the treatment of a CDKL5-related disorder or at least one symptom thereof in a subject; optionally wherein the CDKL5-related disorder is CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome.Attorney Docket No. 14640.0304-00304126. The AAV particle or pharmaceutical composition of claim 125, wherein the subject has, has been diagnosed with having, or is at risk of having the CDKL5-related disorder or at least one symptom thereof; optionally wherein the CDKL5-related disorder is CDD, developmental and epileptic encephalopathy 2, or atypical Rett syndrome.

127. Use of the AAV particle of any one of claims 1-83, the cell of claim 84 or claim 85, or the pharmaceutical composition of claim 93 in the manufacture of a medicament for the treatment of an CDKL5-related disorder or at least one symptom thereof in a subject; optionally wherein the CDKL5-related disorder is CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, or atypical Rett syndrome.

122. The use of claim 121, wherein the subject has, has been diagnosed with having, or is at risk of having the CDKL5-related disorder or at least one symptom thereof; optionally wherein the CDKL5-related disorder is CDD, developmental and epileptic encephalopathy 2, or atypical Rett syndrome.

123. The AAV particle of any one of claims 1-83 or the pharmaceutical composition of claim 93 for use in a method of treating a disorder according to any one of claims 97-124.

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