Compositions and methods for the treatment of disorders associated with syntaxin-binding protein 1 deficiency

Abstract

This disclosure relates to compositions and methods for altering, for example, enhancing, the expression of the STXBP1 protein via delivery using adeno-associated virus (AAV) capsid variants. The compositions and methods of this disclosure are useful in the treatment of subjects diagnosed with or suspected of having STXBP1 encephalopathy or another STXBP1-related disorder.

Description

[Technical Field]

[0001] Sequence List This application is filed together with an electronically formatted sequence listing. The sequence listing file (name: 14640_0085-00304_SL.xml) was created on March 26, 2024, and has a size of 5,536,190 bytes. The electronically formatted sequence listing information is incorporated herein by reference in its entirety.

[0002] field This specification describes compositions and methods relating to adeno-associated virus (AAV) virus particles for delivery of polynucleotides, such as the syntaxin-binding protein-1 (STXBP1) protein ("STXBP1 protein") and peptide ("STXBP1 peptide"), for use in the treatment of STXBP1 encephalopathy, as well as other STXBP1-related disorders, including epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonic epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutations and further mutations by optional selection), Dravet syndrome (not caused by mutations in SCN1A), and Rett syndrome phenotype (not caused by mutations in MECP2 or CDKL5) (collectively, "STXBP1-related disorders"). In some embodiments, the compositions described herein may be used to treat subjects in need, for example, human subjects diagnosed with STXBP1-related disorders or other conditions resulting from deficiencies in the amount and / or function of the STXBP1 protein. [Background technology]

[0003] Syntaxin-binding protein 1 (STXBP1), also known as Munc18-1, is part of the synaptic fusion mechanism that enables vesicles to fuse with the cell membrane. Other names for STXBP1 include P67, DEE4, NSEC1, UNC18, N-Sec1, RBSEC1, unc-18A, and unc18-1. STXBP1 regulates neurotransmitter transmission by interacting with the SNARE complex. The SNARE complex is mainly composed of SNAP-25, vesicle-binding membrane protein, and syntaxin 1. SNAP-25 and syntaxin 1 form a target membrane vesicle protein (T-SNARE) that binds to synaptic vesicle proteins (VAMPs). STXBP1 has a complex arched tertiary structure. The arch contains four closely linked domains, 1, 2, 3a, and 3b. Domains 1 and 3a form an arched gap. STXBP1 primarily regulates vesicle fusion by interacting with syntaxin 1. Domain 3a of STXBP1 is in close contact with the Habc domain of syntaxin-1, and domain 1 of STXBP1 binds to the N-terminal domain of syntaxin-1. STXBP1 regulates vesicle docking and fusion by interacting with the SNARE complex. STXBP1 influences released vesicles and is involved in neurotransmitter transmission. STXBP1 is significantly involved in the initial processes of neurotransmitter release.

[0004] STXBP1 is essential for presynaptic vesicle release. It is rapidly phosphorylated by protein kinase C during neuronal depolarization. STXBP1 is encoded by the STXBP1 gene (Ensembl Gene ID No. ENSG00000136854), located on chromosome 9. It is expressed in the brain and spinal cord and is highly enriched in axons. STXBP1 expression is highest in the retina and cerebellum. STXBP1 is also found outside the brain.

[0005] Mutations in the STXBP1 gene are known to cause disease in human subjects. Abnormal expression of STXBP1 plays a role in the pathogenesis of various neurological disorders, including STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutations and further mutations by random selection), Dravet syndrome (not caused by SCN1A mutations), and Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutations).

[0006] STXBP1 expression is also abnormal in STXBP1 encephalopathy. There are an estimated 750 known cases of STXBP1 encephalopathy worldwide, with an estimated incidence of 3.3 to 3.8 cases per 100,000 births.

[0007] Patients with STXBP1 mutations and / or STXBP1 encephalopathy often present with epilepsy. Some patients with STXBP1 mutations exhibit autistic features, including aggressive behavior, self-injurious behavior, hyperactivity, obsessive-compulsive symptoms, psychiatric disorders, and / or auditory hallucinations. Phenotypes range from severe neonatal epilepsy to infant-onset epilepsy.

[0008] Many patients with STXBP1 mutations also present with non-epileptic movement disorders, including ataxia of the trunk and limbs, generalized tremor, and dystonia. Patients often experience continuous epileptic activity.

[0009] Patients with STXBP1 encephalopathy are dependent on caregivers for life. Furthermore, 40% of patients become unable to walk, and their life expectancy is expected to be drastically shortened to approximately 30 years. STXBP1 encephalopathy is caused by haploinsufficiency. Therefore, the disease can develop if there is a mutation in only a single-function copy of the STXBP1 gene. Disease-causing mutations include missense, nonsense, frameshift, splice site, and deletion of the entire gene. There are approximately 135 known pathogenic variants of the STXBP1 gene.

[0010] Interneurons are more susceptible to haploinsufficiency than excitatory neurons. The study demonstrated that heterozygous STXBP1 knockout mice exhibited moderate seizure phenotypes, as well as impaired glutamate and GABA transmission, increased anxiety, increased aggression, and impaired emotion learning.

[0011] Studies in mice demonstrated that normalization of excitatory synaptic transmission in STXBP1 heterozygous knockout mice reduced aggression. This indicates a therapeutic option for managing aggression in patients with STXBP1 mutations.

[0012] Existing therapies target the symptoms of STXBP1 encephalopathy. Existing first-line treatments include antiepileptic drugs such as levetiracetam and phenobarbital. Existing second-line treatments include additional antiepileptic drugs such as clobazam and topiramate. Existing third-line treatments include additional antiepileptic drugs and / or interventions. Known interventions for existing third-line treatments include adrenocorticotropic hormone, ketogenic diet therapy, and vagus nerve stimulation.

[0013] There are no approved therapies that target the underlying causes of STXBP1 mutations. There is a high unmet need for such treatments, driven by the high seizure rate and shorter life expectancy in patients with STXBP1 mutations.

[0014] Therefore, there has long been a need to develop pharmaceutical compositions and methods that can be delivered to the CNS for the treatment of diseases associated with STXBP1 mutations. In particular, there is a need for therapies that target neurons, including GABAergic and glutamatergic neurons.

[0015] Adeno-associated virus (AAV) has emerged as a viral particle that has been widely studied and utilized for the delivery of therapeutically effective polypeptides into mammalian cells. See, for example, (Non-Patent Document 1) and (Non-Patent Document 2), each of which is incorporated herein by reference in its entirety.

[0016] There is still a need for an effective method of treatment using an AAV capsid variant capable of delivering STXBP1 to target cells or tissues, such as CNS cells or tissues.

Prior Art Documents

Non-Patent Documents

[0017]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

[0018] In some embodiments, the Disclosure provides an AAV capsid variant comprising an amino acid sequence having the following formula:[N1]-[N2]-[N3] (wherein (i) optionally [N1] comprises X1, X2, and X3, where at least one of X1, X2, or X3 is G; (ii) [N2] comprises the amino acid sequence of SPH; and (iii) [N3] comprises X4, X5, and X6, where at least one of X4, X5, or X6 is a basic amino acid); and b) an adeno-associated virus (AAV) particle comprising a viral genome comprising a syntaxin-binding protein 1 (STXBP1) coding sequence. In some embodiments, the amino acid sequence [N1]-[N2]-[N3] is located in the hypervariable loop IV of the AAV capsid variant. In some embodiments, the AAV capsid variant is the AAV9 capsid variant. In some embodiments, [N1] comprises X1, X2, and X3, where at least one of X1, X2, or X3 is G. In some embodiments, [N2]-[N3] comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941).

[0019] In some embodiments, the disclosure provides adeno-associated virus (AAV) particles comprising a viral genome including a syntaxin-binding protein 1 (STXBP1) coding sequence and an AAV9 capsid variant including the amino acid sequence of SPHSKA (SEQ ID NO: 941). In some embodiments, the amino acid sequence of SPHSKA (SEQ ID NO: 941) is located within the hypervariable loop of the AAV9 capsid variant. In some embodiments, the amino acid sequence of SPHSKA (SEQ ID NO: 941) is located immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4 or SEQ ID NO: 36.

[0020] In some embodiments, the AAV9 capsid variant includes one, two, or all of the following amino acids: N at the amino acid position corresponding to position 452 of SEQ ID NO: 4, E at the amino acid position corresponding to position 451, and / or V at the amino acid position corresponding to position 453. In some embodiments, the AAV9 capsid variant includes the amino acid sequence KTENVSGSPHSKAQNQQT (SEQ ID NO: 3272).

[0021] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein having an amino acid sequence having at least 90% identity with SEQ ID NO: 4; (ii) a VP2 protein having an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein having an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 4.

[0022] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein having an amino acid sequence having at least 95% identity with SEQ ID NO: 4; (ii) a VP2 protein having an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein having an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 4.

[0023] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein having an amino acid sequence having at least 99% identity with SEQ ID NO: 4; (ii) a VP2 protein having an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein having an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 4.

[0024] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 4; and / or (iii) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 4.

[0025] In some embodiments, the AAV9 capsid variant (i) contains the amino acid sequence of SPHSKA (SEQ ID NO: 941) (this amino acid sequence is located immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4); (ii) contains E at the amino acid position corresponding to position 451 and V at the amino acid position corresponding to position 453 of SEQ ID NO: 4; and (iii) does not contain any other modifications to wild-type AAV9.

[0026] In some embodiments, the AAV9 capsid variant further includes one, two, or all of the following amino acids: E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452, and / or V at the amino acid position corresponding to position 453. In some embodiments, the AAV9 capsid variant includes the amino acid sequence KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).

[0027] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein having an amino acid sequence having at least 90% identity with SEQ ID NO: 36; (ii) a VP2 protein having an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein having an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 36.

[0028] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein having an amino acid sequence having at least 95% identity with SEQ ID NO: 36; (ii) a VP2 protein having an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein having an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 36.

[0029] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein having an amino acid sequence having at least 99% identity with SEQ ID NO: 36; (ii) a VP2 protein having an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein having an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 36.

[0030] In some embodiments, the AAV9 capsid variant comprises (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 36; and / or (iii) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 36.

[0031] In some embodiments, the AAV9 capsid variant (i) contains the amino acid sequence SPHSKA (SEQ ID NO: 941) (this amino acid sequence is located immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 36); (ii) contains E at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452, and V at the amino acid position corresponding to position 453; and (iii) does not contain any other modifications to wild-type AAV9.

[0032] In some embodiments, the Disclosure provides AAV particles comprising an AAV capsid variant comprising an amino acid sequence having the following formula:[N1]-[N2]-[N3] (wherein [N1]-[N2]-[N3] is located immediately after the position corresponding to amino acid position 452 of SEQ ID NO: 982), wherein the AAV capsid variant comprises an amino acid sequence that is at least 90% identical to, for example, positions 203-742 of SEQ ID NO: 982, for example, 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.

[0033] In some embodiments, [N1] comprises GHD. In some embodiments, [N1] comprises amino acid G at position 453, amino acid H at position 454, and amino acid D at position 455, corresponding to position 453 of SEQ ID NO: 138 or SEQ ID NO: 982. In some embodiments, [N3] comprises KSG.

[0034] In some embodiments, the AAV capsid variant includes (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 982.

[0035] In some embodiments, the AAV capsid variant includes (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 982.

[0036] In some embodiments, the AAV capsid variant includes (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 982.

[0037] In some embodiments, the AAV capsid variant comprises (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 982; (ii) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 982; or (iii) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 982.

[0038] In some embodiments, the disclosure provides AAV particles containing a viral genome encoding the wild-type STXBP1 protein. In some embodiments, the viral genome encodes the human STXBP1 protein. In some embodiments, the STXBP1 protein contains the amino acid sequence of SEQ ID NO: 6413.

[0039] In some embodiments, the STXBP1 coding sequence includes a nucleotide sequence that is at least 90% identical to (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) SEQ ID NO: 6414. In some embodiments, the STXBP1 coding sequence includes a nucleotide sequence that is at least 95% identical to SEQ ID NO: 6414. In some embodiments, the STXBP1 coding sequence includes a nucleotide sequence that is at least 99% identical to SEQ ID NO: 6414. In some embodiments, the STXBP1 coding sequence includes the nucleotide sequence of SEQ ID NO: 6414. In some embodiments, the STXBP1 coding sequence consists of the nucleotide sequence of SEQ ID NO: 6414.

[0040] In some embodiments, the viral genome includes a promoter operably concatenated to the STXBP1 coding sequence. In some embodiments, the promoter is human elongation factor 1α-subunit (EF1α), cytomegalovirus (CMV) early enhancer and / or promoter, chicken β-actin (CBA), CAG, CAG derivatives, β-glucuronidase (GUSB), or ubiquitin C (UBC), neuron-specific enolase (NSE), platelet-derived growth factor (PDGF), platelet-derived growth factor B-chain (PDGF-β), intercellular adhesion molecule 2 (ICAM-2), synapsin (Syn), synapsin 1 (Syn1), methyl-CpG binding protein 2 (MeCP2), Ca2+ / calmodulin-dependent protein kinase II (CaMKII). II) metabotropic glutamate receptor 2 (mGluR2), neurofilament light chain (NFL) or heavy chain (NFH), β-globin minigene nβ2, preproenkephalin (PPE), enkephalin (Enk), and excitatory amino acid transporter 2 (EAAT2), glial fibrillary acidic protein (GFAP), myelin basic protein (MBP), cardiovascular promoters (e.g., αMHC, cTnT, and CMV-MLC2k), hepatic promoters (e.g., hAAT, TBG), skeletal muscle promoters (e.g., desmin, MCK, C512) or their fragments, e.g., shortened forms or functional variants.

[0041] In some embodiments, the viral genome further includes an inverted end (ITR) sequence. In some embodiments, the viral genome includes an ITR sequence located 5' relative to the STXBP1 coding sequence. In some embodiments, the viral genome includes an ITR sequence located 3' relative to the STXBP1 coding sequence. In some embodiments, the viral genome includes an ITR sequence located 5' relative to the STXBP1 coding sequence and an ITR sequence located 3' relative to the STXBP1 coding sequence.

[0042] In some embodiments, the present disclosure provides AAV particles comprising an AAV capsid variant including a viral genome containing the syntaxin-binding protein 1 (STXBP1) coding sequence and (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 4; and / or (iii) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 4.

[0043] In some embodiments, the present disclosure provides AAV particles comprising an AAV capsid variant including a viral genome containing the syntaxin-binding protein 1 (STXBP1) coding sequence and (i) a VP1 protein containing the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 36; and / or (iii) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 36.

[0044] In some embodiments, the disclosure provides cells containing the AAV particles disclosed herein. In some embodiments, the cells are mammalian cells (e.g., HEK293 cells), insect cells (e.g., Sf9 cells), or bacterial cells.

[0045] In some embodiments, the present disclosure provides a method for producing AAV particles as disclosed herein, comprising: (i) providing cells comprising a viral genome comprising a nucleic acid encoding an STXBP1 coding sequence and an AAV capsid variant; and (ii) incubating the cells under conditions suitable for encapsulating the viral genome in the AAV capsid variant; thereby producing AAV particles.

[0046] In some embodiments, the viral genome of the AAV particle comprises the nucleotide sequence of SEQ ID NO: 6414 or a nucleotide sequence that is at least 90% identical thereto (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); and the AAV capsid variant of the AAAV particle comprises (i) the amino acid sequence of SEQ ID NO: 4 or a VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 4 (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% identity); (ii) (iii) A VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 4 (for example, 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% identity); or (iii) a VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 4 (for example, 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% identity).

[0047] In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 4, the amino acid sequence from positions 138 to 742 of SEQ ID NO: 4, and / or the amino acid sequence from positions 203 to 742 of SEQ ID NO: 4.

[0048] In some embodiments, the viral genome of the AAV particle comprises the nucleotide sequence of SEQ ID NO: 6414 or a nucleotide sequence that is at least 90% identical thereto (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); and the AAV capsid variant of the AAV particle comprises (i) the amino acid sequence of SEQ ID NO: 36 or a VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 36 (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% identity); and (ii) the sequence (iii) A VP2 protein comprising an amino acid sequence having at least 90% identity (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%) with the amino acid sequence at positions 138-742 of sequence number 36; or a VP3 protein comprising an amino acid sequence having at least 90% identity (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%) with the amino acid sequence at positions 203-742 of sequence number 36.

[0049] In some embodiments, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 36, the amino acid sequence from positions 138 to 742 of SEQ ID NO: 36, and / or the amino acid sequence from positions 203 to 742 of SEQ ID NO: 36.

[0050] In some embodiments, the viral genome of the AAV particle includes the nucleotide sequence of SEQ ID NO: 6414 or a nucleotide sequence that is at least 90% identical thereto (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); and the AAV capsid variant of the AAAV particle includes (i) the amino acid sequence of SEQ ID NO: 982 or a VP1 protein having at least 90% identity with SEQ ID NO: 982 (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% identity); and (ii) the sequence (iii) A VP2 protein comprising an amino acid sequence having at least 90% identity (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%) with the amino acid sequence at positions 138-742 of sequence number 982; or a VP3 protein comprising an amino acid sequence having at least 90% identity (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%) with the amino acid sequence at positions 203-742 of sequence number 982.

[0051] In some embodiments, the AAV capsid variant includes the amino acid sequence of SEQ ID NO: 982, the amino acid sequence from positions 138 to 742 of SEQ ID NO: 982, and / or the amino acid sequence from positions 203 to 742 of SEQ ID NO: 982.

[0052] In some embodiments, the preparation method further includes introducing a first nucleic acid molecule containing the viral genome into cells prior to step (i). In some embodiments, the cells contain a second nucleic acid molecule encoding an AAV capsid variant. In some embodiments, the preparation method further includes introducing the second nucleic acid molecule into the cells prior to step (i).

[0053] In some embodiments, the cells include mammalian cells (e.g., HEK293 cells), insect cells (e.g., Sf9 cells), or bacterial cells. In some embodiments, the present disclosure provides pharmaceutical compositions comprising AAV particles and pharmaceutically acceptable excipients as disclosed herein.

[0054] In some embodiments, the present disclosure provides a method for delivering AAV particles encoding the STXBP1 protein to a target, comprising administering an effective amount of the pharmaceutical composition or AAV particles disclosed herein to the target.

[0055] In some embodiments, the Disclosure provides a method for treating a subject having or diagnosed with an STXBP1-related disorder, comprising administering to the subject an effective amount of a pharmaceutical composition or AAV particles disclosed herein. In some embodiments, the STXBP1-related disorder is an STXBP1-related neurodegenerative disorder or neuromuscular disorder. In some embodiments, the STXBP1-related neurodegenerative or neuromuscular disorder is an STXBP1 encephalopathy, an epileptic encephalopathy, Ohtahara syndrome, a developmental encephalopathy, West syndrome, an early myoclonic epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with an STXBP1 mutation and further mutations by choice), Dravet syndrome (not caused by an SCN1A mutation), or Rett syndrome phenotype (not caused by an MECP2 or CDKL5 mutation).

[0056] In some embodiments, the Disclosure provides a method for treating a subject having or diagnosed with STXBP1 encephalopathy, comprising administering to the subject an effective amount of a pharmaceutical composition or AAV particles disclosed herein.

[0057] In some embodiments, the subjects have one or more mutations in the STXBP1 gene. In some embodiments, the subjects have a reduced level of STXBP1 activity compared to a reference level in subjects without STXBP1-related impairment.

[0058] In some embodiments, the treatment results in prevention of disability progression in the subject. In some embodiments, the treatment results in recovery from the disability. In some embodiments, the treatment results in changes in one or more biomarkers of the disability. In some embodiments, one or more biomarkers include STXBP1 activity or neurofilament light chains.

[0059] In some embodiments, the treatment results in the recovery of at least one symptom of the disorder. In some embodiments, the at least one symptom includes epilepsy, autistic features, ataxia, generalized tremor, dystonia, or a combination thereof.

[0060] In some embodiments, the subject is a human. In some embodiments, AAV particles are delivered to cells, tissues, or regions of the CNS, such as regions of the brain or spinal cord, such as parenchyma, cortex, substantia nigra, caudate nucleus-cerebellum, striatum, corpus callosum, cerebellum, brainstem, caudate nucleus-putamen, thalamus, superior colliculus, spinal cord, or a combination thereof, and / or neurons, such as GABAergic neurons, glutamatergic neurons, or a combination thereof.

[0061] In any one of the preceding embodiments, the method of delivery or treatment further includes evaluating, for example, measuring, the level of STXBP1 expression in a subject, for example, in the cells, tissues, or body fluids of the subject, e.g., STXBP1 gene expression, STXBP1 mRNA expression, and / or STXBP1 protein expression. In some embodiments, the level of STXBP1 protein expression is measured by ELISA, Western blotting, or immunohistochemical assay.

[0062] In some embodiments, the level of STXBP1 expression is evaluated before and after administration of AAV particles, and optionally, the level of STXBP1 expression before treatment is compared to the level of STXBP1 expression after administration. In some embodiments, the level of STXBP1 expression is evaluated in cells or tissues of the central nervous system (e.g., parenchyma) from the subject. In some embodiments, the level of STXBP1 protein expression in the subject after administration is increased compared to the level of STXBP1 protein expression in the subject before administration.

[0063] In some embodiments, administration of the pharmaceutical compositions or AAV particles described herein results in (i) STXBP1 activity in the cells, tissues (e.g., cells or tissues of the CNS, e.g., cortex, striatum, thalamus, cerebellum, and / or brainstem) and / or body fluids (e.g., CSF and / or serum) of the subject compared to STXBP1 activity in the subject before administration; (ii) levels of viral genome (VG) per cell in the subject's CNS tissue (e.g., cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord) compared to levels of VG per cell in the subject's peripheral tissue; and / or (iii) increased STXBP1 mRNA expression in the cells or tissues (e.g., cells or tissues of the CNS, e.g., cortex, thalamus, and / or brainstem) compared to STXBP1 mRNA expression in the subject before administration.

[0064] In some embodiments, the treatment method further comprises administering to the subject an additional agent suitable for the treatment or prevention of STXBP1-related disorder. In some embodiments, the additional agent includes one or more antiepileptic drugs (e.g., levetiracetam, phenobarbital, clobazam, topiramate), adrenocorticotropic hormone, or a combination thereof. In some embodiments, the method further comprises administering to the subject an immunosuppressant. In some embodiments, the immunosuppressant includes corticosteroids (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxychloroquine.

[0065] In some embodiments, the Disclosure provides pharmaceutical compositions or AAV particles described herein for use in the treatment of STXBP1-related disorders. In some embodiments, STXBP1-related disorders include STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutations and further mutations by optional selection), Dravet syndrome (not caused by SCN1A mutations), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutations).

[0066] In some embodiments, this disclosure provides the use of the pharmaceutical compositions or AAV particles described herein in the manufacture of a medicament for the treatment of STXBP1-related disorders. In some embodiments, STXBP1-related disorders include STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutations and further mutations by optional selection), Dravet syndrome (not caused by SCN1A mutations), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutations).

[0067] Enumerated embodiments 1. Adeno-associated virus (AAV) particles comprising a viral genome containing an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is given by the following formula: [N1]-[N2]-[N3](wherein, (i) By choice, [N1] includes X1, X2, and X3, where at least one of X1, X2, or X3 is G; (ii)[N2] contains the amino acid sequence of SPH; and (iii)[N3] comprises X4, X5, and X6, where at least one of X4, X5, or X6 is a basic amino acid, e.g., K or R. Adeno-associated virus (AAV) particles containing an amino acid sequence having the following characteristics.

[0068] 2. AAV particle of Embodiment 1, wherein X4, X5, or both of [N3] are K. 3. AAV particles of Embodiment 1 or 2, wherein X4, X5, or X6 of [N3] is R.

[0069] 4. (a) X4 of [N3] is K, S, A, V, T, G, F, W, V, N, or R; (b) X5 of [N3] is S, K, T, F, I, L, Y, H, M, or R; and / or (c) X6 of [N3] is G, A, R, M, I, N, T, Y, D, P, V, L, E, W, N, Q, K, or S; Optionally, an AAV particle of any one of Embodiments 1 to 3, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (c), e.g., a conservative substitution.

[0070] 5. AAV particles of any one of Embodiments 1 to 4, wherein [N3] includes SK, KA, KS, AR, RM, VK, AS, SR, VK, KR, KK, KN, VR, RS, RK, KT, TS, KF, FG, KI, IG, KL, LG, TT, TY, KY, YG, KD, KP, TR, RG, VR, GA, SL, SS, FL, WK, SA, RA, LR, KW, RR, GK, TK, NK, AK, KV, KG, KH, KM, TG, SE, SV, SW, SN, HG, SQ, LW, MG, MA, or SG.

[0071] 6. An AAV particle of any one of Embodiments 1 to 5, wherein [N3] is or comprises SKA, KSG, ARM, VKS, ASR, VKI, KKN, VRM, RKA, KTS, KFG, KIG, KLG, KTT, KTY, KYG, SKD, SKP, TRG, VRG, KRG, GAR, KSA, KSR, SKL, SRA, SKR, SLR, SRG, SSR, FLR, SKW, SKS, WKA, VRR, SKV, SKT, SKG, GKA, TKA, NKA, SKL, SKN, AKA, KTG, KSL, KSE, KSV, KSW, KSN, KHG, KSQ, KSK, KLW, WKG, KMG, KMA, or RSG.

[0072] 7. [N2]-[N3] are SPHSK (sequence number 4701), SPHKS (sequence number 4704), SPHAR (sequence number 4705), SPHVK (sequence number 4706), SPHAS (sequence number 4707), SPHKK (sequence number 4708), SPHVR (sequence number 4709), SPHRK (sequence number 4710), SPHKT (sequence number 4711), SPHKF (sequence number 4712), SPHKI (sequence number 4713), SPHKL (sequence number 4714), SPHKY (sequence number 4715), SPHTR (sequence number 4716), SPHKR (distribution AAV particles from any one of Embodiments 1 to 6, comprising: (sequence number 4717), SPHGA (sequence number 4718), SPHSR (sequence number 4719), SPHSL (sequence number 4720), SPHSS (sequence number 4721), SPHFL (sequence number 4722), SPHWK (sequence number 4723), SPHGK (sequence number 4724), SPHTK (sequence number 4725), SPHNK (sequence number 4726), SPHAK (sequence number 4727), SPHKH (sequence number 4728), SPHKM (sequence number 4729), or SPHRS (sequence number 4730).

[0073] 8. [N2]-[N3] is (i) SPHSKA (SEQ ID NO: 941), SPHKSG (SEQ ID NO: 946), SPHARM (SEQ ID NO: 947), SPHVKS (SEQ ID NO: 948), SPHASR (SEQ ID NO: 949), SPHVKI (SEQ ID NO: 950), SPHKKN (SEQ ID NO: 954), SPHVRM (SEQ ID NO: 955), SPHRKA (SEQ ID NO: 956), SPHKFG (SEQ ID NO: 957), SPHKIG (SEQ ID NO: 958), SPHKLG (SEQ ID NO: 959), SPHKTS (SEQ ID NO: 963), SPHKTT (SEQ ID NO: 964), SPHKTY ( SEQ ID NO: 965), SPHKYG (SEQ ID NO: 966), SPHSKD (SEQ ID NO: 967), SPHSKP (SEQ ID NO: 968), SPHTRG (SEQ ID NO: 972), SPHVRG (SEQ ID NO: 973), SPHKRG (SEQ ID NO: 974), SPHGAR (SEQ ID NO: 975), SPHKSA (SEQ ID NO: 977), SPHKSR (SEQ ID NO: 951), SPHSKL (SEQ ID NO: 960), SPHSRA (SEQ ID NO: 969), SPHSKR (SEQ ID NO: 978), SPHSLR (SEQ ID NO: 952), SPHSRG (SEQ ID NO: 961), SP HSSR (SEQ ID NO: 970), SPHFLR (SEQ ID NO: 979), SPHSKW (SEQ ID NO: 953), SPHSKS (SEQ ID NO: 962), SPHWKA (SEQ ID NO: 971), SPHVRR (SEQ ID NO: 980), SPHSKT (SEQ ID NO: 4731), SPHSKG (SEQ ID NO: 4732), SPHGKA (SEQ ID NO: 4733), SPHNKA (SEQ ID NO: 4734), SPHSKN (SEQ ID NO: 4735), SPHAKA (SEQ ID NO: 4736), SPHSKV (SEQ ID NO: 4737), SPHKTG (SEQ ID NO: 4738), SPHT KA (sequence code 4739), SPHKSL (sequence code 4740), SPHKSE (sequence code 4741), SPHKSV (sequence code 4742), SPHKSW (sequence code 4743), SPHKSN (sequence code 4744), SPHKHG (sequence code 4745), SPHKSQ (sequence code 4746), SPHKSK (sequence code 4747), SPHKLW (sequence code 4748), SPHWKG (sequence code 4749), SPHKMG (sequence code 4750), SPHKMA (sequence code 4751), or SPHRSG (sequence code 976); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of Embodiments 1 to 7.

[0074] 9. An AAV particle of any one of Embodiments 1 to 8, wherein the AAV capsid variant is numbered according to one of Sequence IDs 36-59, 138, 981, or 982, comprising an amino acid other than G at position 453 (e.g., V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C), an amino acid other than S at position 454 (e.g., V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q), and / or an amino acid other than G at position 455 (e.g., C, L, D, E, Y, H, V, A, N, P, or S).

[0075] 10. An AAV particle of any one of Embodiments 1 to 8, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or 981, and comprises amino acid G at position 453, amino acid S at position 454, and amino acid G at position 455.

[0076] 11. An AAV particle of any one of Embodiments 1 to 9, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or 982, and comprises amino acid G at position 453, amino acid H at position 454, and amino acid D at position 455.

[0077] 12. An AAV particle of any one of embodiments 1 to 11, wherein [N1] comprises X1, X2, and X3, and at least one of X1, X2, or X3 is G. 13. (a) X1 in [N1] is G, V, R, D, E, M, T, I, S, A, N, L, K, H, P, W, or C; (b) X2 of [N1] is S, V, L, N, D, H, R, P, G, T, I, A, E, Y, M, or Q; and / or (c) X3 of [N1] is G, C, L, D, E, Y, H, V, A, N, P, or S; Optionally, one AAV particle of any of Embodiments 1 to 12, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (c), e.g., a conservative substitution.

[0078] 14. AAV particles of any one of Embodiments 1 to 13, wherein [N1] comprises GS, SG, GH, HD, GQ, QD, VS, CS, GR, RG, QS, SH, MS, RN, TS, IS, GP, ES, SS, GN, AS, NS, LS, GG, KS, GT, PS, RS, GI, WS, DS, ID, GL, DA, DG, ME, EN, KN, KE, AI, NG, PG, TG, SV, IG, LG, AG, EG, SA, YD, HE, HG, RD, ND, PD, MG, QV, DD, HN, HP, GY, GM, GD, or HS.

[0079] 15. AAV particles of any one of Embodiments 1 to 14, wherein [N1] is or comprises GSG, GHD, GQD, VSG, CSG, GRG, CSH, GQS, GSH, RVG, GSC, GLL, GDD, GHE, GNY, MSG, RNG, TSG, ISG, GPG, ESG, SSG, GNG, ASG, NSG, LSG, GGG, KSG, HSG, GTG, PSG, GSV, RSG, GIG, WSG, DSG, IDG, GLG, DAG, DGG, MEG, ENG, GSA, KNG, KEG, AIG, GYD, GHG, GRD, GND, GPD, GMG, GQV, GHN, GHP, or GHS.

[0080] 16. [N1]-[N2] is, (i)SGSPH (SEQ ID NO: 4752), HDSPH (SEQ ID NO: 4703), QDSPH (SEQ ID NO: 4753), RGSPH (SEQ ID NO: 4754), SHSPH (SEQ ID NO: 4755), QSSPH (SEQ ID NO: 4756), DDSPH (SEQ ID NO: 4757), HESPH (SEQ ID NO: 4758), NYSPH (SEQ ID NO: 4759), VGSPH (SEQ ID NO: 4760), SCSPH (SEQ ID NO: 4761), LLSPH (SEQ ID NO: 4762), NGSPH (SEQ ID NO: 4763), PGSPH (SEQ ID NO: 4764), GGSPH (SEQ ID NO: 4765), TGSPH (SEQ ID NO: 4766), SVSPH (SEQ ID NO: 4752), Column number 4767), IGSPH (sequence number 4768), DGSPH (sequence number 4769), LGSPH (sequence number 4770), AGSPH (sequence number 4771), EGSPH (sequence number 4772), SASPH (sequence number 4773), YDSPH (sequence number 4774), HGSPH (sequence number 4775), RDSPH (sequence number 4776), NDSPH (sequence number 4777), PDSPH (sequence number 4778), MGSPH (sequence number 4779), QVSPH (sequence number 4780), HNSPH (sequence number 4781), HPSPH (sequence number 4782), or HSSPH (sequence number 4783); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, or four amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle comprising any one of embodiments 1 to 15.

[0081] 17. [N1]-[N2] is (i) GSGSPH (SEQ ID NO: 4695), GHDSPH (SEQ ID NO: 4784), GQDSPH (SEQ ID NO: 4785), VSGSPH (SEQ ID NO: 4786), CSGSPH (SEQ ID NO: 4787), GRGSPH (SEQ ID NO: 4788), CSHSPH (SEQ ID NO: 4789), GQSSPH (SEQ ID NO: 4790), GSHSPH (SEQ ID NO: 4791), GDDSPH (SEQ ID NO: 4792), GHESPH (SEQ ID NO: 4793), GNYSPH (SEQ ID NO: 4794), RVGSPH (SEQ ID NO: 4795), GSCSPH (SEQ ID NO: 4796) ), GLLSPH (sequence number 4797), MSGSPH (sequence number 4798), RNGSPH (sequence number 4799), TSGSPH (sequence number 4800), ISGSPH (sequence number 4801), GPGSPH (sequence number 4802), ESGSPH (sequence number 4803), SSGSPH (sequence number 4804), GNGSPH (sequence number 4805), ASGSPH (sequence number 4806), NSGSPH (sequence number 4807), LSGSPH (sequence number 4808), GGGSPH (sequence number 4809), KSGSPH (sequence number 4810) HSGSPH (sequence number 4811), GTGSPH (sequence number 4812), PSGSPH (sequence number 4813), GSVSPH (sequence number 4814), RSGSPH (sequence number 4815), GIGSPH (sequence number 4816), WSGSPH (sequence number 4817), DSGSPH (sequence number 4818), IDGSPH (sequence number 4819), GLGSPH (sequence number 4820), DAGSPH (sequence number 4821), DGGSPH (sequence number 4822), MEGSPH (sequence number 4823), ENGSPH (sequence number 4824), GSASPH (sequence number 4825), KNGSPH (sequence number 4826), KEGSPH (sequence number 4827), AIGSPH (sequence number 4828), GYDSPH (sequence number 4829), GHGSPH (sequence number 4830), GRDSPH (sequence number 4831), GNDSPH (sequence number 4832), GPDSPH (sequence number 4833), GMGSPH (sequence number 4834), GQVSPH (sequence number 4835), GHNSPH (sequence number 4836), GHPSPH (sequence number 4837), or GHSSPH (sequence number 4838); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 1 to 16.

[0082] 18. [N1]-[N2]-[N3] is (i)SGSPHSK (SEQ ID NO: 4839), HDSPHKS (SEQ ID NO: 4840), SGSPHAR (SEQ ID NO: 4841), SGSPHVK (SEQ ID NO: 4842), QDSPHKS (SEQ ID NO: 4843), SGSPHKK (SEQ ID NO: 4844), SGSPHVR (SEQ ID NO: 4845), SGSPHAS (SEQ ID NO: 4846), SGSPHRK (SEQ ID NO: 4847), SGSPHKT (SEQ ID NO: 4848), SHSPHKS (SEQ ID NO: 4849), QSSPHRS (SEQ ID NO: 4850), RGSPHAS (SEQ ID NO: 4851), RGSPHSK (SEQ ID NO: 4849) SGSPHKF (SEQ ID NO: 4852), SGSPHKI (SEQ ID NO: 4854), SGSPHKL (SEQ ID NO: 4855), SGSPHKY (SEQ ID NO: 4856), SGSPHTR (SEQ ID NO: 4857), SHSPHKR (SEQ ID NO: 4858), SGSPHGA (SEQ ID NO: 4859), HDSPHKR (SEQ ID NO: 4860), DDSPHKS (SEQ ID NO: 4861), HESPHKS (SEQ ID NO: 4862), NYSPHKI (SEQ ID NO: 4863), SGSPHSR (SEQ ID NO: 4864), SGSPHSL (SEQ ID NO: 4865), SGSPHSS (SEQ ID NO: 4866), VGSPHSK (SEQ ID NO: 4867), SCSPHRK (SEQ ID NO: 4868), SGSPHFL (SEQ ID NO: 4869), LLSPHWK (SEQ ID NO: 4870), NGSPHSK (SEQ ID NO: 4871), PGSPHSK (SEQ ID NO: 4872), GGSPHSK (SEQ ID NO: 4873), TGSPHSK (SEQ ID NO: 4874), SVSPHGK (SEQ ID NO: 4875), SGSPHTK (SEQ ID NO: 4876), IGSPHSK (SEQ ID NO: 4877), DGSPHSK (SEQ ID NO: 4878), SGSPHNK (SEQ ID NO: 4879), LGS PHSK (SEQ ID NO: 4880), AGSPHSK (SEQ ID NO: 4881), EGSPHSK (SEQ ID NO: 4882), SASPHSK (SEQ ID NO: 4883), SGSPHAK (SEQ ID NO: 4884), HDSPHKI (SEQ ID NO: 4885), YDSPHKS (SEQ ID NO: 4886), HDSPHKT (SEQ ID NO: 4887), RGSPHKR (SEQ ID NO: 4888), HGSPHSK (SEQ ID NO: 4889), RDSPHKS (SEQ ID NO: 4890), NDSPHKS (SEQ ID NO: 4891), QDSPHKI (SEQ ID NO: 4892), PDSPHKI (SEQ ID NO: 4893),PDSPHKS (sequence code 4894), MGSPHSK (sequence code 4895), HDSPHKH (sequence code 4896), QVSPHKS (sequence code 4897), HNSPHKS (sequence code 4898), NGSPHKR (sequence code 4899), HDSPHKY (sequence code 4900), NDSPHKI (sequence code 4901), HDSPHKL (sequence code 4902), HPSPHWK (sequence code 4903), HDSPHKM (sequence code 4904), or HSSPHRS (sequence code 4905); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, five, or six amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle comprising any one of embodiments 1 to 17.

[0083] 19. [N1]-[N2]-[N3] is (i) GSGSPHSKA (SEQ ID NO: 4697), GHDSPHKSG (SEQ ID NO: 4698), GSGSPHARM (SEQ ID NO: 4906), GSGSPHVKS (SEQ ID NO: 4907), GQDSPHKSG (SEQ ID NO: 4908), GSGSPHASR (SEQ ID NO: 4909), GSGSPHVKI (SEQ ID NO: 4910), GSGSPHKKN (SEQ ID NO: 4911), GSGSPHVRM (SEQ ID NO: 4912), VSGSPHSKA (SEQ ID NO: 4913), CSGSPHSKA (SEQ ID NO: 4914), GSGSPHRKA (SEQ ID NO: 4915), CS GSPHKTS (SEQ ID NO: 4916), CSHSPHKSG (SEQ ID NO: 4917), GQSSPHRSG (SEQ ID NO: 4918), GRGSPHASR (SEQ ID NO: 4919), GRGSPHSKA (SEQ ID NO: 4920), GSGSPHKFG (SEQ ID NO: 4921), GSGSPHKIG (SEQ ID NO: 4922), GSGSPHKLG (SEQ ID NO: 4923), GSGSPHKTS (SEQ ID NO: 4924), GSGSPHKTT (SEQ ID NO: 4925), GSGSPHKTY (SEQ ID NO: 4926), GSGSPHKYG (SEQ ID NO: 4927), GSGSPHSK D (SEQ ID NO: 4928), GSGSPHSKP (SEQ ID NO: 4929), GSGSPHTRG (SEQ ID NO: 4930), GSGSPHVRG (SEQ ID NO: 4931), GSHSPHKRG (SEQ ID NO: 4932), GSHSPHKSG (SEQ ID NO: 4933), VSGSPHASR (SEQ ID NO: 4934), VSGSPHGAR (SEQ ID NO: 4935), VGSPHKFG (SEQ ID NO: 4936), GHDSPHKRG (SEQ ID NO: 4937), GDDSPHKSG (SEQ ID NO: 4938), GHESPHKSA (SEQ ID NO: 4939), GHDSPHKSA (SEQ ID NO: 4938) 4940), GNYSPHKIG (SEQ ID NO: 4941), GHDSPHKSR (SEQ ID NO: 4942), GSGSPHSKL (SEQ ID NO: 4943), GSGSPHSRA (SEQ ID NO: 4944), GSGSPHSKR (SEQ ID NO: 4945), GSGSPHSLR (SEQ ID NO: 4946), GSGSPHSRG (SEQ ID NO: 4947), GSGSPHSSR (SEQ ID NO: 4948), RVGSPHSKA (SEQ ID NO: 4949), GSCSPHRKA (SEQ ID NO: 4950), GSGSPHFLR (SEQ ID NO: 4951), GSGSPHSKW (SEQ ID NO: 4952),GSGSPHSKS (SEQ ID NO: 4953), GLLSPHWKA (SEQ ID NO: 4954), GSGSPHVRR (SEQ ID NO: 4955), GSGSPHSKV (SEQ ID NO: 4956), MSGSPHSKA (SEQ ID NO: 4957), RNGSPHSKA (SEQ ID NO: 4958), TSGSPHSKA (SEQ ID NO: 4959), ISGSPHSKA (SEQ ID NO: 4960), GPGSPHSKA (SEQ ID NO: 4961), GSGSPHSKT (SEQ ID NO: 4962), ESGSPHSKA (SEQ ID NO: 4963), SSGSPHSKA (SEQ ID NO: 4964), GNGSPHSKA ( SEQ ID NO: 4965), ASGSPHSKA (SEQ ID NO: 4966), NSGSPHSKA (SEQ ID NO: 4967), LSGSPHSKA (SEQ ID NO: 4968), GGGSPHSKA (SEQ ID NO: 4969), KSGSPHSKA (SEQ ID NO: 4970), GGGSPHSKS (SEQ ID NO: 4971), GSGSPHSKG (SEQ ID NO: 4972), HSGSPHSKA (SEQ ID NO: 4973), GTGSPHSKA (SEQ ID NO: 4974), PSGSPHSKA (SEQ ID NO: 4975), GSVSPHGKA (SEQ ID NO: 4976), RSGSPHSKA (SEQ ID NO: 4977), GSGSPHTKA (SEQ ID NO: 4978), GIGSPHSKA (SEQ ID NO: 4979), WSGSPHSKA (SEQ ID NO: 4980), DSGSPHSKA (SEQ ID NO: 4981), IDGSPHSKA (SEQ ID NO: 4982), GSGSPHNKA (SEQ ID NO: 4983), GLGSPHSKS (SEQ ID NO: 4984), DAGSPHSKA (SEQ ID NO: 4985), DGGSPHSKA (SEQ ID NO: 4986), MEGSPHSKA (SEQ ID NO: 4987), ENGSPHSKA (SEQ ID NO: 4988), GSASPHSKA (SEQ ID NO: 4989), GNGSPHSKS ( SEQ ID NO: 4990), KNGSPHSKA (SEQ ID NO: 4991), KEGSPHSKA (SEQ ID NO: 4992), AIGSPHSKA (SEQ ID NO: 4993), GSGSPHSKN (SEQ ID NO: 4994), GSGSPHAKA (SEQ ID NO: 4995), GHDSPHKIG (SEQ ID NO: 4996), GYDSPHKSG (SEQ ID NO: 4997), GHESPHKSG (SEQ ID NO: 4998), GHDSPHKTG (SEQ ID NO: 4999), GRGSPHKRG (SEQ ID NO: 5000), GQDSPHKSG (SEQ ID NO: 4908), GHDSPHKSL (SEQ ID NO: 5001),GHGSPHSKA (SEQ ID NO: 5002), GHDSPHKSE (SEQ ID NO: 5003), VSGSPHSKA (SEQ ID NO: 4913), GRDSPHKSG (SEQ ID NO: 5004), GNDSPHKSV (SEQ ID NO: 5005), GQDSPHKIG (SEQ ID NO: 5006), GHDSPHKSV (SEQ ID NO: 5007), GPDSPHKIG (SEQ ID NO: 5008), GPDSPHKSG (SEQ ID NO: 5009), GHDSPHKSW (SEQ ID NO: 5010), GHDSPHKSN (SEQ ID NO: 5011), GMGSPHSKT (SEQ ID NO: 5012), GHDSPHKHG (SEQ ID NO: 5013), G QVSPHKSG (SEQ ID NO: 5014), GDDSPHKSV (SEQ ID NO: 5015), GHNSPHKSG (SEQ ID NO: 5016), GNGSPHKRG (SEQ ID NO: 5017), GHDSPHKYG (SEQ ID NO: 5018), GHDSPHKSQ (SEQ ID NO: 5019), GNDSPHKIG (SEQ ID NO: 5020), GHDSPHKSK (SEQ ID NO: 5021), GHDSPHKLW (SEQ ID NO: 5022), GHPSPHWKG (SEQ ID NO: 5023), GHDSPHKMG (SEQ ID NO: 5024), GHDSPHKMA (SEQ ID NO: 5025), or GHSSPHRSG (SEQ ID NO: 5026); (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, or 8 amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 1 to 18.

[0084] 20. An AAV particle from any one of Embodiments 1 to 19, wherein [N3] comprises SK, KA, KS, or SG. 21. An AAV particle from any one of Embodiments 1 to 20, wherein [N3] is or contains SKA, KSG, or KYG.

[0085] 22. An AAV particle from any one of Embodiments 1 to 21, wherein [N2]-[N3] includes SPHSK (SEQ ID NO: 4701), SPHKS (SEQ ID NO: 4704), or SPHKY (SEQ ID NO: 4715).

[0086] 23. An AAV particle from any one of Embodiments 1 to 22, wherein [N2]-[N3] is SPHSKA (SEQ ID NO: 941) or contains the same. 24. An AAV particle from any one of Embodiments 1 to 22, wherein [N2]-[N3] is SPHKSG (SEQ ID NO: 946) or contains the same.

[0087] 25. An AAV particle from any one of Embodiments 1 to 22, wherein [N2]-[N3] is SPHKYG (SEQ ID NO: 966) or contains the same. 26. An AAV particle from any one of Embodiments 1 to 25, wherein [N1] comprises GS, SG, GH, or HD.

[0088] 27. An AAV particle from any one of Embodiments 1 to 26, wherein [N1] is or contains GSG. 28. An AAV particle from any one of Embodiments 1 to 26, wherein [N1] is or contains a GHD.

[0089] 29. AAV particles from any one of Embodiments 1-23 or 26-27, wherein [N1]-[N2]-[N3] include SGSPHSK (SEQ ID NO: 4839). 30. A single AAV particle from any one of Embodiments 1 to 22, 24, 26, or 28, wherein [N1]-[N2]-[N3] includes HDSPHKS (SEQ ID NO: 4840).

[0090] 31. AAV particles from any one of Embodiments 1-22 or 25-27, wherein [N1]-[N2]-[N3] contain SGSPHKYG (SEQ ID NO: 5027). 32. AAV particles from any one of Embodiments 1-8, 10, 12-23, 26-27, or 29, wherein [N1]-[N2]-[N3] is or contains GSGSPHSKA (SEQ ID NO: 4697).

[0091] 33. A single AAV particle from any one of Embodiments 1-9, 11-22, 24, 26, 28, or 30, wherein [N1]-[N2]-[N3] is or contains GHDSPHKSG (SEQ ID NO: 4698).

[0092] 34. One AAV particle from any of the following groups: 1-8, 10, 12-22, 25-27, or 31, in which [N1]-[N2]-[N3] is GSGSPHKYG (Sequence ID 4927) or contains it.

[0093] 35. Any one of Embodiments 1 to 34, wherein [N1]-[N2]-[N3] replace positions 453-455, which are numbered according to Sequence ID No. 138. 36. An AAV particle of any one of Embodiments 1 to 35, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, comprising an amino acid other than Q at position 456 (e.g., W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F), an amino acid other than N at position 457 (e.g., Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L), an amino acid other than Q at position 458 (e.g., G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y), and / or an amino acid other than Q at position 459 (e.g., H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V).

[0094] 37. The AAV capsid variant is numbered according to SEQ ID NOs. 981, 982, 36, 37, 39, 40, 42-46, 48, 49, 50, 52, 53, 56, or 57, with an amino acid other than Q at position 462 (e.g., W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F), and an amino acid other than N at position 463 (e.g., Y, C, K, T, H, R, D, V, S, P, G, An AAV particle comprising any one of Embodiments 1 to 36, comprising W, E, F, A, I, M, Q, or L, an amino acid other than Q at position 464 (e.g., G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y), and / or an amino acid other than Q at position 465 (e.g., H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V).

[0095] 38. AAV capsid variants, (a) Amino acid Q at position 456, amino acid N at position 457, amino acid Q at position 458, and / or amino acid Q at position 459, numbered according to Sequence ID No. 138; or (b) Numbered according to Sequence IDs 981, 982, 36, 37, 39, 40, 42-46, 48, 49, 50, 52, 53, 56, or 57, with amino acid Q at position 462, amino acid N at position 463, amino acid Q at position 464, and / or amino acid Q at position 465. An AAV particle comprising any one of embodiments 1 to 37.

[0096] 39. The AAV capsid variant further comprises [N4], and [N4] comprises X7, X8, X9, X10, and (a) X7 is Q, W, K, R, G, L, V, S, P, H, K, I, M, A, E, or F; (b) X8 is N, Y, C, K, T, H, R, D, V, S, P, G, W, E, F, A, I, M, Q, or L; (c) X9 is Q, G, K, H, R, T, L, D, A, P, I, F, V, M, W, Y, S, E, N, or Y; and (d) X10 is Q, H, L, R, W, K, A, P, E, M, I, S, G, N, Y, C, V, T, D, or V; Optionally, one AAV particle from any of Embodiments 1 to 38, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (d), e.g., a conservative substitution.

[0097] 40. (a) X7 of [N4] is either Q or R; (b) X8 of [N4] is either N or R; (c) X9 in [N4] is Q or R; and (d) X10 of [N4] is Q, L, or R. AAV particles of Embodiment 39.

[0098] 41. [N4] ga, (i) QNQQ (SEQ ID NO: 5028), WNQQ (SEQ ID NO: 5029), QYYV (SEQ ID NO: 5030), RRQQ (SEQ ID NO: 5031), GCGQ (SEQ ID NO: 5032), LRQQ (SEQ ID NO: 5033), RNQQ (SEQ ID NO: 5034), VNQQ (SEQ ID NO: 5035), FRLQ (SEQ ID NO: 5036), FNQQ (SEQ ID NO: 5037), LLQQ (SEQ ID NO: 5038), SNQQ (SEQ ID NO: 5039), RLQQ (SEQ ID NO: 5040), LNQQ (SEQ ID NO: 5041), QRKL (SEQ ID NO: 5042), LRRQ (SEQ ID NO: 5043), QRLR ( SEQ ID NOs: 5044, QRRL (SEQ ID NOs: 5045), RRLQ (SEQ ID NOs: 5046), RLRQ (SEQ ID NOs: 5047), SKRQ (SEQ ID NOs: 5048), QLYR (SEQ ID NOs: 5049), QLTV (SEQ ID NOs: 5050), QNKQ (SEQ ID NOs: 5051), KNQQ (SEQ ID NOs: 5052), QKQQ (SEQ ID NOs: 5053), QTQQ (SEQ ID NOs: 5054), QNHQ (SEQ ID NOs: 5055), QHQQ (SEQ ID NOs: 5056), QNQH (SEQ ID NOs: 5057), QHRQ (SEQ ID NOs: 5058), LTQQ (SEQ ID NOs: 5059), QNQW (SEQ ID NOs: 5060) QNTH (sequence number 5061), RRRQ (sequence number 5062), QYQQ (sequence number 5063), QNDQ (sequence number 5064), QNRH (sequence number 5065), RDQQ (sequence number 5066), PNLQ (sequence number 5067), HVRQ (sequence number 5068), PNQH (sequence number 5069), HNQQ (sequence number 5070), QSQQ (sequence number 5071), QPAK (sequence number 5072), QNLA (sequence number 5073), QNQL (sequence number 5074), QGQQ (sequence number 5075), LNRQ (sequence number 5076), QNPP (distribution) Column number 5077), QNLQ (sequence number 5078), QDQE (sequence number 5079), QDQQ (sequence number 5080), HWQQ (sequence number 5081), PNQQ (sequence number 5082), PEQQ (sequence number 5083), QRTM (sequence number 5084), LHQH (sequence number 5085), QHRI (sequence number 5086), QYIH (sequence number 5087), QKFE (sequence number 5088), QFPS (sequence number 5089), QNPL (sequence number 5090), QAIK (sequence number 5091), QNRQ (sequence number 5092), QYQH (sequence number 5093),QNPQ (sequence number 5094), QHQL (sequence number 5095), QSPP (sequence number 5096), QAKL (sequence number 5097), KSQQ (sequence number 5098), QDRP (sequence number 5099), QNLG (sequence number 5100), QAFH (sequence number 5101), QNAQ (sequence number 5102), HNQL (sequence number 5103), QKLN (sequence number 5104), QNVQ (sequence number 5105), QAQQ (sequence number 5106), QTPP (sequence number 5107), QPPA (sequence number 5108), QERP (sequence number 5109), QDLQ (array (Sequence number 5110), QAMH (Sequence number 5111), QHPS (Sequence number 5112), PGLQ (Sequence number 5113), QGIR (Sequence number 5114), QAPA (Sequence number 5115), QIPP (Sequence number 5116), QTQL (Sequence number 5117), QAPS (Sequence number 5118), QNTY (Sequence number 5119), QDKQ (Sequence number 5120), QNHL (Sequence number 5121), QIGM (Sequence number 5122), LNKQ (Sequence number 5123), PNQL (Sequence number 5124), QLQQ (Sequence number 5125), QRMS (Sequence number 5126), QGIL (SEQ ID NO: 5127), QDRQ (SEQ ID NO: 5128), RDWQ (SEQ ID NO: 5129), QERS (SEQ ID NO: 5130), QNYQ (SEQ ID NO: 5131), QRTC (SEQ ID NO: 5132), QIGH (SEQ ID NO: 5133), QGAI (SEQ ID NO: 5134), QVPP (SEQ ID NO: 5135), QVQQ (SEQ ID NO: 5136), LMRQ (SEQ ID NO: 5137), QYSV (SEQ ID NO: 5138), QAIT (SEQ ID NO: 5139), QKTL (SEQ ID NO: 5140), QLHH (SEQ ID NO: 5141), QNII (SEQ ID NO: 5142), QGHH (SEQ ID NO: 5142) (Sequence number 5143), QSKV (Sequence number 5144), QLPS (Sequence number 5145), IGKQ (Sequence number 5146), QAIH (Sequence number 5147), QHGL (Sequence number 5148), QFMC (Sequence number 5149), QNQM (Sequence number 5150), QHLQ (Sequence number 5151), QPAR (Sequence number 5152), QSLQ (Sequence number 5153), QSQL (Sequence number 5154), HSQQ (Sequence number 5155), QMPS (Sequence number 5156), QGSL (Sequence number 5157), QVPA (Sequence number 5158), HYQQ (Sequence number 5159),QVPS (SEQ code 5160), RGEQ (SEQ code 5161), PGQQ (SEQ code 5162), LEQQ (SEQ code 5163), QNQS (SEQ code 5164), QKVI (SEQ code 5165), QNND (SEQ code 5166), QSVH (SEQ code 5167), QPLG (SEQ code 5168), HNQE (SEQ code 5169), QIQQ (SEQ code 5170), QVRN (SEQ code 5171), PSNQ (SEQ code 5172), QVGH (SEQ code 5173), QRDI (SEQ code 5174), QMPN (SEQ code 5175), RGLQ (SEQ code) (Sequence number 5176), PSLQ (Sequence number 5177), QRDQ (Sequence number 5178), QAKG (Sequence number 5179), QSAH (Sequence number 5180), QSTM (Sequence number 5181), QREM (Sequence number 5182), QYRA (Sequence number 5183), QRQQ (Sequence number 5184), QWQQ (Sequence number 5185), QRMN (Sequence number 5186), GDSQ (Sequence number 5187), QKIS (Sequence number 5188), PSMQ (Sequence number 5189), SPRQ (Sequence number 5190), MEQQ (Sequence number 5191), QYQN (Sequence number 5192), QIRQ (sequence number 5193), QSVQ (sequence number 5194), RSQQ (sequence number 5195), QNKL (sequence number 5196), QIQH (sequence number 5197), PRQQ (sequence number 5198), HTQQ (sequence number 5199), QRQH (sequence number 5200), RNQE (sequence number 5201), QSKQ (sequence number 5202), QNQP (sequence number 5203), QSPQ (sequence number 5204), QTRQ (sequence number 5205), QNLH (sequence number 5206), QNQE (sequence number 5207), LNQP (sequence number 5208), QNQD (sequence number) (Sequence number 5209), QNLL (Sequence number 5210), QLVI (Sequence number 5211), RTQE (Sequence number 5212), QTHQ (Sequence number 5213), QDQH (Sequence number 5214), QSQH (Sequence number 5215), VRQQ (Sequence number 5216), AWQQ (Sequence number 5217), QSVP (Sequence number 5218), QNIQ (Sequence number 5219), LDQQ (Sequence number 5220), PDQQ (Sequence number 5221), ESQQ (Sequence number 5222), QRQL (Sequence number 5223), QIIV (Sequence number 5224), QKQS (Sequence number 5225),QSHQ (sequence number 5226), QFVV (sequence number 5227), QSQP (sequence number 5228), QNEQ (sequence number 5229), INQQ (sequence number 5230), RNRQ (sequence number 5231), RDQK (sequence number 5232), QWKR (sequence number 5233), ENRQ (sequence number 5234), QTQP (sequence number 5235), QKQL (sequence number 5236), RNQL (sequence number 5237), ISIQ (sequence number 5238), QTVC (sequence number 5239), QQIM (sequence number 5240), LNHQ (sequence number 5241), QNQA (sequence number 5242), QMIH (sequence number 5243), RNHQ (sequence number 5244), or QKMN (sequence number 5245); (ii) Any portion of the amino acid sequence in (i), for example, any two or three amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 39 or 40, which are or include such particles.

[0099] 42. [N1]-[N2]-[N3]-[N4] is (i) Any amino acid sequence from sequence numbers 1800 to 2241; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 39 to 41.

[0100] 43. Any one of the AAV particles from Embodiments 39 to 42, wherein [N1]-[N2]-[N3]-[N4] is or contains GSGSPHSKAQNQQ (Sequence ID 1801).

[0101] 44. Any one of the AAV particles from Embodiments 39 to 42, wherein [N1]-[N2]-[N3]-[N4] is GHDSPHKSGQNQQ (SEQ ID NO: 1800) or contains the same.

[0102] 45. An AAV particle from any one of Embodiments 39 to 42, wherein [N1]-[N2]-[N3]-[N4] is or contains GSGSPHKYGQNQQT (SEQ ID NO: 910).

[0103] 46. ​​An AAV particle of any one of Embodiments 1 to 45, wherein the AAV capsid variant is numbered according to one of Sequence IDs 36-59, 138, 981, or 982, comprising an amino acid other than T at position 450 (e.g., S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G), an amino acid other than I at position 451 (e.g., M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L), and / or an amino acid other than N at position 452 (e.g., M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S).

[0104] 47. An AAV particle of any one of Embodiments 1 to 46, wherein the AAV capsid variant is numbered according to any one of Sequence ID No. 138, 981, or 982, and comprises amino acid T at position 450, amino acid I at position 451, and / or amino acid N at position 452.

[0105] 48. The AAV capsid variant further contains [N0], where [N0] is X A , X B and X C including and (a)X A However, it is T, S, Y, M, A, C, I, R, L, D, F, V, Q, N, H, E, or G; (b)X B but is I, M, P, E, N, D, S, A, T, G, Q, F, V, L, C, H, R, W, or L; and (c)X C is N, M, E, G, Y, W, T, I, Q, F, V, A, L, I, P, K, R, H, S, D, or S; and Optionally, one AAV particle from any of Embodiments 1 to 47, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (c), e.g., a conservative substitution.

[0106] 49. [N0] is TIN, SMN, TIM, YLS, GLS, MPE, MEG, MEY, AEW, CEW, ANN, IPE, ADM, IEY, ADY, IET, MEW, CEY, RI N, MEI, LEY, ADW, IEI, DIM, FEQ, MEF, CDQ, LPE, IEN, MES, AEI, VEY, IIN, TSN, IEV, MEM, AEV, MDA, VE W, AEQ, LEW, MEL, MET, MEA, IES, MEV, CEI, ATN, MDG, QEV, ADQ, NMN, IEM, ISN, TGN, QQQ, HDW, IEG, TII, TFP, TEK, EIN, TVN, TFN, SIN, TER, TSY, ELH, AIN, SVN, TDN, TFH, TVH, TEN, TSS, TID, TCN, NIN, TE H, AEM, AIK, TDK, TFK, SDQ, TEI, NTN, TET, SIK, TEL, TEA, TAN, TIY, TFS, TES, TTN, TED, TNN, EVH, TI S, TVR, TDR, TIK, NHI, TIP, ESD, TDL, TVP, TVI, AEH, NCL, TVK, NAD, TIT, NCV, TIR, NAL, VIN, TIQ, TE The AAV particle of embodiment 48 is or comprises a dipeptide of F, TRE, QGE, SEK, NVN, GGE, EFV, SDK, TEQ, EVQ, TEY, NCW, TDV, SDI, NSI, NSL, EVV, TEP, SEL, TWQ, TEV, AVN, GVL, TLN, TEG, TRD, NAI, AEN, AET, ETA, NNL, or any of them.

[0107] 50. [N0]-[N1]-[N2]-[N3]-[N4] is (i) Any one amino acid sequence from sequence numbers 2242 to 2886; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 48 or 49, which are or include such particles.

[0108] 51. Any one of the AAV particles from Embodiments 48 to 50, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains TINGSGSPHSKAQNQQ (Sequence ID 2242).

[0109] 52. Any one of the AAV particles from Embodiments 48 to 50, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains TINGHDSPHKSGQNQQ (Sequence ID 2243).

[0110] 53. Any one of the AAV particles from Embodiments 48 to 52, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains TINGSGSPHKYGQNQQT (Sequence ID 5246).

[0111] 54. One AAV particle from any of Embodiments 1 to 53, wherein [N1]-[N2]-[N3] is present in Loop IV. 55. One of the AAV particles from any of embodiments 48 to 54, wherein [N0] and [N4] are present in loop IV.

[0112] 56. An AAV particle from any one of embodiments 48 to 55, wherein [N0] is located immediately after position 449, which is numbered according to sequence number 138. 57. An AAV particle of any one of embodiments 48 to 56, wherein [N0] is located immediately after position 449, numbered according to any one of sequence numbers 36 to 59, 981, or 982.

[0113] 58. An AAV particle of any one of embodiments 48 to 57, wherein [N0] replaces positions 450, 451, and 452 (e.g., T450, I451, and N452) numbered according to sequence number 138.

[0114] 59. An AAV particle of any one of Embodiments 48 to 58, wherein [N0] replaces positions 450 to 452 (e.g., T450, I451, and N452) numbered according to any one of Sequence IDs 36 to 59, 981, or 982.

[0115] 60. An AAV particle from any one of Embodiments 48 to 59, wherein [N0] corresponds to position 450 to 452 of any one of Sequence IDs 36 to 59, 138, 981, or 982. 61. Any one of the AAV particles from Embodiments 48 to 60, wherein [N0] is located immediately after position 449, which is numbered according to Sequence ID No. 138, and [N0] replaces positions 450 to 452 (e.g., T450, I451, and N452).

[0116] 62. An AAV particle of any one of embodiments 48 to 61, wherein [N0] is located immediately after position 449, which is numbered according to any one of sequence numbers 36 to 59, 981, or 982, and [N0] replaces positions 450 to 452 (e.g., T450, I451, and N452).

[0117] 63. Any one of Embodiments 1 to 62, wherein [N1] is located immediately after position 452, which is numbered according to the amino acid sequence of Sequence ID No. 138. 64. An AAV particle from any one of Embodiments 1 to 63, wherein [N1] is located immediately after position 452, as numbered according to Sequence ID No. 981 or 982.

[0118] 65. An AAV particle from any one of Embodiments 1 to 61, wherein [N1] replaces positions 453 to 455 (e.g., G453, S454, and G455) numbered according to Sequence ID No. 138.

[0119] 66. An AAV particle of any one of Embodiments 1 to 64, wherein [N1] replaces position 453 (e.g., G453) as numbered according to Sequence ID No. 138. 67. An AAV particle of any one of Embodiments 1 to 65, wherein [N1] replaces positions 453 to 455 (e.g., G453, S454, and G455) numbered according to Sequence ID No. 981.

[0120] 68. An AAV particle of any one of Embodiments 1 to 65 or 67, wherein [N1] replaces positions 453 to 455, which are numbered according to Sequence ID No. 982. 69. An AAV particle of any one of embodiments 1 to 65, 67, or 68, wherein [N1] is located immediately after position 452, which is numbered according to sequence number 138, and [N1] replaces positions 453 to 455 (e.g., G453, S454, and G455).

[0121] 70. An AAV particle of any one of embodiments 1 to 64 or 66, wherein [N1] is located immediately after position 452, which is numbered according to sequence number 138, and [N1] replaces position 453 (e.g., G453).

[0122] 71. An AAV particle of any one of embodiments 1 to 64, 66, or 70, wherein [N1] is located immediately after position 452, which is numbered according to sequence numbers 4, 36, 981, or 982, and [N1] replaces positions 453 to 455.

[0123] 72. An AAV particle from any one of Embodiments 1 to 71, wherein [N1] corresponds to positions 453 to 455, numbered according to any one of Sequence IDs 4, 36 to 59, 981, or 982.

[0124] 73. An AAV particle of any one of Embodiments 1 to 72, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, 981, or 982, and contains an amino acid other than S at position 454 and / or an amino acid other than G at position 455.

[0125] 74. An AAV particle of any one of Embodiments 1 to 73, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or 982, and comprises amino acid H at position 454 and amino acid D at position 455.

[0126] 75. Any one of Embodiments 1 to 74, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, and includes substitution at position 454 (e.g., S454H) and / or substitution at position 455 (e.g., G455D).

[0127] 76. Any one of the AAV particles from Embodiments 1 to 75, wherein the AAV capsid variant is numbered according to SEQ ID NO: 138, contains amino acid H at position 454 and amino acid D at position 455, and further contains the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately after position 455.

[0128] 77. Any one of the AAV particles from Embodiments 1 to 76, wherein the AAV capsid variant comprises amino acid H at position 454 and amino acid D at position 455, as numbered according to Sequence ID No. 982.

[0129] 78. Any one of the AAV particles from Embodiments 1 to 77, wherein the AAV capsid variant is numbered according to SEQ ID NO: 982, contains amino acid H at position 454 and amino acid D at position 455, and further contains the amino acid sequence SPHSKA (SEQ ID NO: 941) immediately after position 455.

[0130] 79. An AAV particle of any one of Embodiments 1 to 72, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, and comprises amino acid S at position 454 and amino acid G at position 455.

[0131] 80. An AAV particle of any one of Embodiments 1 to 72 or 79, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, contains amino acid S at position 454 and amino acid G at position 455, and further contains the amino acid sequence SPHSKA (Sequence ID No. 941) immediately after position 455.

[0132] 81. An AAV particle of any one of Embodiments 1 to 72, 79, or 80, wherein the AAV capsid variant is numbered according to Sequence ID No. 981 and contains amino acid S at position 454 and amino acid G at position 455.

[0133] 82. An AAV particle of any one of Embodiments 1-72 or 79-81, wherein the AAV capsid variant is numbered according to Sequence ID No. 981, contains amino acid S at position 454 and amino acid G at position 455, and further contains the amino acid sequence SPHSKA (Sequence ID No. 941) immediately after position 455.

[0134] 83. An AAV particle from any one of Embodiments 1 to 82, wherein [N2] is located immediately after position 455, as numbered according to Sequence ID No. 138. 84. An AAV particle from any one of Embodiments 1 to 83, wherein [N2] corresponds to positions 456-458 of Sequence ID No. 981 or 982 (e.g., S456, P457, H458).

[0135] 85. An AAV particle from any one of Embodiments 1 to 83, wherein [N2] corresponds to position 456 to 458 (e.g., S456, P457, H458) of any one of Sequence ID No. 4 or 36 to 59.

[0136] 86. An AAV particle from any one of Embodiments 1 to 85, wherein [N2]-[N3] is located immediately after position 455, as numbered according to Sequence ID No. 138. 87. An AAV particle from any one of Embodiments 1 to 86, wherein [N2] is located immediately after position 455, as numbered according to Sequence ID No. 4, 36, 981, or 982.

[0137] 88. Any one of the AAV particles from Embodiments 1 to 87, wherein [N2]-[N3] is located immediately after position 455, as numbered according to Sequence ID No. 4, 36, 981, or 982. 89. An AAV particle from any one of Embodiments 1 to 88, wherein [N2]-[N3] corresponds to positions 456-461 of Sequence ID No. 981 (e.g., S456, P457, H458, S459, K460, A461).

[0138] 90. An AAV particle from any one of Embodiments 1 to 88, wherein [N2]-[N3] corresponds to positions 456-461 of Sequence ID No. 982 (e.g., S456, P457, H458, K459, S460, G461).

[0139] 91. An AAV particle from any one of Embodiments 1 to 90, wherein [N2] is located immediately after [N1]. 92. Any one of the AAV particles of Embodiments 1 to 64, 66, 70, or 71, wherein [N3] is located immediately after [N2] and replaces positions 454 and 455 (e.g., S454 and G455) numbered according to Sequence ID No. 138.

[0140] 93. One AAV particle of any one of Embodiments 1-1-64, 66, 70, 71, or 92, wherein [N3] is located immediately after [N1]-[N2] and replaces positions 454 and 455 (e.g., S454 and G455) numbered according to Sequence ID No. 138.

[0141] 94. An AAV particle from any one of embodiments 39 to 93, wherein [N4] is located immediately after the 455th position, as numbered according to sequence number 138. 95. Any one of the AAV particles of Embodiments 39 to 94, wherein [N4] replaces positions 456 to 459 (e.g., Q456, N457, Q458, and Q459) numbered according to Sequence ID No. 138.

[0142] 96. An AAV particle from any one of Embodiments 39 to 95, wherein [N4] corresponds to positions 462 to 465 of Sequence ID No. 4, 36, 981, or 982 (e.g., Q462, N463, Q464, Q465).

[0143] 97. Any one of the AAV particles from Embodiments 39 to 96, wherein [N2]-[N3]-[N4] replace positions 456-459 (e.g., Q456, N457, Q458, and Q459) numbered according to Sequence ID No. 138.

[0144] 98. Any one of the AAV particles of Embodiments 39 to 97, wherein [N2]-[N3]-[N4] is located immediately after position 455, which is numbered according to Sequence ID No. 138, and [N2]-[N3]-[N4] replaces positions 456-459 (e.g., Q456, N457, Q458, and Q459).

[0145] 99. One of the AAV particles from any one of Embodiments 39 to 98, wherein [N2]-[N3]-[N4] corresponds to positions 456-465 of Sequence ID No. 981 (e.g., S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465).

[0146] 100. One of the AAV particles from any one of Embodiments 39 to 98, wherein [N2]-[N3]-[N4] corresponds to positions 456-465 of Sequence ID No. 982 (e.g., S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465).

[0147] 101. An AAV particle from any one of Embodiments 39 to 98, wherein [N2]-[N3]-[N4] corresponds to positions 456-465 of one of Sequence ID No. 4 or 36-59. 102. Any one of the AAV particles from Embodiments 39 to 101, wherein [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, and Q459) numbered according to Sequence ID No. 138.

[0148] 103. Any one of the AAV particles of Embodiments 39 to 102, wherein [N1]-[N2]-[N3]-[N4] is located immediately after position 452, which is numbered according to Sequence ID No. 138, and [N1]-[N2]-[N3]-[N4] replaces positions 453-459 (e.g., G453, S454, G455, Q456, N457, Q458, and Q459).

[0149] 104. One AAV particle from any one of embodiments 39-99, 102, or 103, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-465 of sequence number 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465).

[0150] 105. One AAV particle from any one of embodiments 39-98, 100, 102, or 103, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-465 of sequence number 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465).

[0151] 106. An AAV particle in any one of embodiments 39-98, 102, or 103, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-465 of any one of sequence numbers 4 or 36-59.

[0152] 107. One AAV particle from any of Embodiments 1-99 or 102-104, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 of Sequence ID No. 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461).

[0153] 108. One AAV particle from any one of Embodiments 1-98, 100, 102, 103, or 105, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 of Sequence ID No. 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461).

[0154] 109. An AAV particle from any one of embodiments 39-98, 102, 103, or 106, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 of any one of sequence numbers 36-59.

[0155] 110. Any one of the AAV particles from Embodiments 48 to 109, wherein [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-459 (e.g., T450, I451, N452, G453, S454, G455, Q456, N457, Q458, and Q459) numbered according to Sequence ID No. 138.

[0156] 111. Any one of the AAV particles of Embodiments 48 to 110, wherein [N0]-[N1]-[N2]-[N3]-[N4] is located immediately after position 449, numbered according to Sequence ID No. 138, and [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 450-459 (e.g., T450, I451, N452, G453, S454, G455, Q456, N457, Q458, and Q459).

[0157] 112. One AAV particle from any one of embodiments 48-99, 102-104, or 106, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 450-465 of sequence number 981 (e.g., T450, I451, N452, G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465).

[0158] 113. One of the AAV particles from embodiments 48-98, 100, 102, 103, 105, or 108, where [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 450-465 of sequence number 982 (e.g., T450, I451, N452, G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465).

[0159] 114. An AAV particle of any one of embodiments 48-98, 102, 103, 106, or 109, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to position 450-465 of the sequence number or any one of 36-59.

[0160] 115. Any one of the AAV particles of Embodiments 39 to 114, wherein [N4] replaces positions 462 to 465 (e.g., Q462, N463, Q464, and Q465) numbered according to Sequence ID No. 4, 36, 981, or 982.

[0161] 116. Any one of the AAV particles of Embodiments 39 to 115, wherein [N2]-[N3]-[N4] replace positions 462-465 (e.g., Q462, N463, Q464, and Q465) numbered according to Sequence ID No. 4, 36, 981, or 982.

[0162] 117. Any one of the AAV particles of Embodiments 39 to 116, wherein [N2]-[N3]-[N4] is located immediately after position 455, which is numbered according to Sequence ID No. 4, 36, 981, or 982, and [N2]-[N3]-[N4] replaces positions 462-465 (e.g., Q462, N463, Q464, and Q465).

[0163] 118. One AAV particle from any of Embodiments 1 to 117, wherein [N3] is located immediately after [N2]. 119. Any one of the AAV particles from Embodiments 1 to 118, wherein the AAV capsid variant comprises [N2]-[N3] from the N-terminus to the C-terminus.

[0164] 120. Any one of the AAV particles from Embodiments 1 to 119, wherein the AAV capsid variant comprises [N1]-[N2]-[N3] from the N-terminus to the C-terminus. 121. Any one of the AAV particles from Embodiments 48 to 120, wherein the AAV capsid variant comprises [N0]-[N1]-[N2]-[N3] from the N-terminus to the C-terminus.

[0165] 122. Any one of the AAV particles from Embodiments 39 to 121, wherein the AAV capsid variant comprises [N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus. 123. Any one of the AAV particles from Embodiments 48 to 122, wherein the AAV capsid variant comprises [N0]-[N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0166] 124. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant contains an amino acid other than T at position 460, numbered according to Sequence ID No. 138 (e.g., N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S).

[0167] 125. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant contains amino acids N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S at position 460, as numbered according to Sequence ID No. 138.

[0168] 126. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant contains an amino acid other than T at position 466, numbered according to any one of SEQ ID NOs. 36-59, 981, or 982 (e.g., N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S).

[0169] 127. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant contains amino acids N, I, C, H, R, L, D, Y, A, M, Q, I, E, K, P, G, or S at position 466, numbered according to any one of Sequence IDs 36-59, 981, or 982.

[0170] 128. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant contains an amino acid other than K (e.g., E, N, or T) at position 449, numbered according to any one of SEQ ID NOs. 36-59, 138, 981, or 982.

[0171] 129. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant contains amino E, N, or T at position 449, numbered according to any one of SEQ ID NOs: 36-59, 138, 981, or 982.

[0172] 130. Adeno-associated virus (AAV) particles comprising a viral genome containing an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant comprises [A][B] (SEQ ID NO: 4694), (i)[A] contains the amino acid sequence of GSGSPH (SEQ ID NO: 4695); and (ii)[B] includes X1 X2 X3 X4 X5 X6 X7, (a) X1 is S, C, F, or V; (b) X2 is K, L, R, I, E, Y, V, or S; (c) X3 is A, R, L, G, I, Y, S, F, or W; (d) X4 is W, Q, R, G, L, V, S, or F; (e) X5 is N, Y, R, C, K, or L; (f) X6 is Q, G, K, R, T, L, or Y; and (g) X7 is Q, L, R, or V; By choice, adeno-associated virus (AAV) particles in which the AAV capsid variant includes an amino acid modification, e.g., a conservative substitution, of any of the aforementioned amino acids in (a) to (g).

[0173] 131. (a) X1 is S; (b) X2 is either K or L; (c) X3 is A, R, or L; (d) X4 is Q or R; (e) X5 is N or R; (f) X6 is Q or R; and (g) X7 is Q, L, or R. AAV particles of Embodiment 130.

[0174] 132. [B] is, (i) SLLWNQQ (SEQ ID NO: 5247), SKAQYYV (SEQ ID NO: 5248), SKLRRQQ (SEQ ID NO: 5249), SIWQNQQ (SEQ ID NO: 5250), SKAGCGQ (SEQ ID NO: 5251), SRAQNQQ (SEQ ID NO: 5252), SKRLRQQ (SEQ ID NO: 5253), SLRRNQQ (SEQ ID NO: 5254), SRGRNQQ (SEQ ID NO: 5255), SEIVNQ Q (sequence number 5256), SSRRNQQ (sequence number 5257), CLLQNQQ (sequence number 5258), SKAFRLQ (sequence number 5259), CLAQNQQ (sequence number 5260), FLRQNQQ (sequence number 5261), SLRFNQQ (sequence number 5262), SYLRNQQ (sequence number 5263), CSLQNQQ (sequence number 5264), VLWQNQQ (sequence number 5265) ), SKWLLQQ (sequence number 5266), SLWSNQQ (sequence number 5267), SKRRLQQ (sequence number 5268), SVYLNQQ (sequence number 5269), SLWLNQQ (sequence number 5270), SKAQRKL (sequence number 5271), SKALRRQ (sequence number 5272), SKAQRLR (sequence number 5273), SKAQNQQ (sequence number 5274), SKAQRRL ( SEQ ID NOs: 5275), SKARRQQ (SEQ ID NOs: 5276), SKARRQQ (SEQ ID NOs: 5277), SKSRRQQ (SEQ ID NOs: 5278), SKARRQQ (SEQ ID NOs: 5279), SKASKRQ (SEQ ID NOs: 5280), VRRQNQQ (SEQ ID NOs: 5281), SKAQLYR (SEQ ID NOs: 5282), SLFRNQQ (SEQ ID NOs: 5283), SKAQLTV (SEQ ID NOs: 5284); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, five, or six amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of embodiment 130 or 131, including the above.

[0175] 133. [A][B] (i) GSGSPHSLLWNQQ (SEQ ID NO: 5285), GSGSPHSKAQYYV (SEQ ID NO: 2060), GSGSPHSKLRRQQ (SEQ ID NO: 2061), GSGSPHSIWQNQQ (SEQ ID NO: 5286), GSGSPHSKAGCGQ (SEQ ID NO: 2062), GSGSPHSRAQNQQ (SEQ ID NO: 2063), GSGSPHSKRLRQQ (SEQ ID NO: 2064), GSGSPHSLRRNQQ (SEQ ID NO: 2065), GSGSPHSRGRNQQ (SEQ ID NO: 2066), GGSPHSEI VNQQ (SEQ ID NO: 5287), GSGSPHSSRRNQQ (SEQ ID NO: 2067), GSGSPHCLLQNQQ (SEQ ID NO: 5288), GSGSPHSKAFRLQ (SEQ ID NO: 2068), GSGSPHCLAQNQQ (SEQ ID NO: 5289), GSGSPHFLRQNQQ (SEQ ID NO: 2070), GSGSPHSLRFNQQ (SEQ ID NO: 2071), GSGSPHSYLRNQQ (SEQ ID NO: 5290), GSGSPHCSLQNQQ (SEQ ID NO: 5291), GSGSPHVLWQNQQ (SEQ ID NO: 5292 ), GSGSPHSKWLLQQ (Sequence ID 2072), GSGSPHSLWSNQQ (Sequence ID 5293), GSGSPHSKRRLQQ (Sequence ID 2073), GSGSPHSVYLNQQ (Sequence ID 5294), GSGSPHSLWLNQQ (Sequence ID 5295), GSGSPHSKAQRKL (Sequence ID 2074), GSGSPHSKALRRQ (Sequence ID 2075), GSGSPHSKAQRLR (Sequence ID 2076), GSGSPHSKAQNQQ (Sequence ID 1801), GSGSPHSKAQR RL (sequence number 2077), GSGSPHSKARRQQ (sequence number 2078), GSGSPHSKARRLQ (sequence number 2079), GSGSPHSKSRRQQ (sequence number 2080), GSGSPHSKARLRQ (sequence number 2082), GSGSPHSKASKRQ (sequence number 2083), GSGSPHVRRQNQQ (sequence number 2084), GSGSPHSKAQLYR (sequence number 2085), GSGSPHSLFRNQQ (sequence number 5296), GSGSPHSKAQLTV (sequence number 2086); (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle comprising any one of embodiments 130 to 132.

[0176] 134. Any one of the AAV particles of Embodiments 130 to 133, wherein the AAV capsid variant further comprises one, two, or all of the following, numbered according to Sequence ID No. 138: an amino acid other than T at position 450 (e.g., S, Y, or G), an amino acid other than I at position 451 (e.g., M or L), and / or an amino acid other than N at position 452 (e.g., S).

[0177] 135. Any one of the AAV particles of Embodiments 130 to 134, wherein the AAV capsid variant further comprises S at position 450 and M at position 451, numbered according to Sequence ID No. 138.

[0178] 136. Any one of the AAV particles of Embodiments 130 to 134, wherein the AAV capsid variant further comprises Y at position 450, L at position 451, and S at position 452, numbered according to Sequence ID No. 138.

[0179] 137. Any one of the AAV particles of Embodiments 130 to 134, wherein the AAV capsid variant further comprises G at position 450, L at position 451, and S at position 452, as numbered according to Sequence ID No. 138.

[0180] 138. A single AAV particle from any one of embodiments 130 to 137, wherein [A] and [B] are present in loop IV. 139. An AAV particle from any one of embodiments 130 to 138, wherein [A] is located immediately after position 452, numbered according to sequence number 138.

[0181] 140. An AAV particle of any one of embodiments 130 to 139, wherein [A] replaces positions 453 to 455 (e.g., G453, S454, G455) numbered according to sequence number 138.

[0182] 141. Any one of the AAV particles of Embodiments 130 to 140, wherein [A] is located immediately after position 452, numbered according to Sequence ID No. 138, and [A] replaces positions 453 to 455 (e.g., G453, S454, G455).

[0183] 142. An AAV particle from any one of embodiments 130 to 141, wherein [B] is located immediately after [A]. 143. Any one of the AAV particles from Embodiments 130 to 142, wherein [B] replaces positions 456 to 459 (e.g., Q456, N457, Q458, Q459) numbered according to Sequence ID No. 138.

[0184] 144. Any one of the AAV particles from Embodiments 130 to 143, wherein [A][B] replaces positions 453 to 459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459) numbered according to Sequence ID No. 138.

[0185] 145. Any one of the AAV particles of Embodiments 130 to 144, wherein [A][B] is located immediately after position 452, numbered according to Sequence ID No. 138, and [A][B] replaces positions 453 to 459 (e.g., G453, S454, G455, Q456, N457, Q458, Q459).

[0186] 146. An AAV particle of any one of embodiments 130 to 145, wherein the AAV capsid variant comprises [A] and [B] from the N-terminus to the C-terminus. 147. Adeno-associated virus (AAV) particles comprising a viral genome containing an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant comprises [A][B] (SEQ ID NO: 4699), (i)[A] includes X1 X2 X3 X4 X5 X6, (a) X1 is T, M, A, C, I, R, L, D, F, V, Q, N, or H; (b) X2 is I, P, E, N, D, S, A, T, M, or Q; (c) X3 is N, E, G, Y, W, M, T, I, K, Q, F, S, V, A, or L; (d) X4 is G, D, R, or E; (e) X5 is H, Q, N, or D; (f) X6 is either D or R; By choice, the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (f), e.g., a conservative substitution; and (ii)[B] contains SPHKSG (sequence number 946), Adeno-associated virus (AAV) particles.

[0187] 148. (a) X1 is T, M, A, or I; (b) X2 is E, I, or D; (c) X3 is N, Q, Y, I, M, or V; (d) X4 is G; (e) X5 is H; and (f) X6 is D, AAV particles of Embodiment 147.

[0188] 149. [A] is, (i) TINGHD (SEQ ID NO: 5297), MPEGHD (SEQ ID NO: 5298), MEGGHD (SEQ ID NO: 5299), MEYGHD (SEQ ID NO: 5300), AEWGHD (SEQ ID NO: 5301), CEWGHD (SEQ ID NO: 5302), ANNGQD (SEQ ID NO: 5303), IPEGHD (SEQ ID NO: 5304), ADMGHD (SEQ ID NO: 5305), IEYGHD (SEQ ID NO: 5306), ADYGHD (SEQ ID NO: 5307), IETGHD (SEQ ID NO: 5308), MEWGHD (SEQ ID NO: 5309), CEYGHD (SEQ ID NO: 5310), R INGHD (SEQ ID NO: 5311), MEIGHD (SEQ ID NO: 5312), LEYGHD (SEQ ID NO: 5313), ADWGHD (SEQ ID NO: 5314), IEIGHD (SEQ ID NO: 5315), TIKDND (SEQ ID NO: 5316), DIMGHD (SEQ ID NO: 5317), FEQGHD (SEQ ID NO: 5318), MEFGHD (SEQ ID NO: 5319), CDQGHD (SEQ ID NO: 5320), LPEGHD (SEQ ID NO: 5321), IENGHD (SEQ ID NO: 5322), MESGHD (SEQ ID NO: 5323), AEIGHD (SEQ ID NO: 5324), VEYGHD (SEQ ID NO: 5325), TSNGDD (SEQ ID NO: 5326), IEGGHD (SEQ ID NO: 5327), MEMGHD (SEQ ID NO: 5328), AEVGHD (SEQ ID NO: 5329), MDAGHD (SEQ ID NO: 5330), VEWGHD (SEQ ID NO: 5331), AEQGHD (SEQ ID NO: 5332), LEWGHD (SEQ ID NO: 5333), MELGHD (SEQ ID NO: 5334), METGHD (SEQ ID NO: 5335), MEAGHD (SEQ ID NO: 5336), TINRQR (SEQ ID NO: 5337), IESGHD (SEQ ID NO: 5338), TAKDHD (SEQ ID NO: 5335) 5339), MEVGHD (SEQ ID NO: 5340), CEIGHD (SEQ ID NO: 5341), ATNGHD (SEQ ID NO: 5342), MDGGHD (SEQ ID NO: 5343), QEVGHD (SEQ ID NO: 5344), ADQGHD (SEQ ID NO: 5345), NMNGHD (SEQ ID NO: 5346), TPWEHD (SEQ ID NO: 5347), IEMGHD (SEQ ID NO: 5348), TANEHD (SEQ ID NO: 5349), QQQGHD (SEQ ID NO: 5350), TPQDHD (SEQ ID NO: 5351), HDWGHD (SEQ ID NO: 5352), IEGGHD (SEQ ID NO: 5353) (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of embodiment 147 or 148, including the above.

[0189] 150. [A][B] (i)TINGHDSPHKR (SEQ ID NO: 5354), MPEGHDSPHKS (SEQ ID NO: 5355), MEGGHDSPHKS (SEQ ID NO: 5356), MEYGHDSPHKS (SEQ ID NO: 5357), AEWGHDSPHKS (SEQ ID NO: 5358), CEWGHDSPHKS (SEQ ID NO: 5359), ANNGQDSPHKS (SEQ ID NO: 5360), IPEGHDSPHKS (SEQ ID NO: 5361), ADMGHDSPHKS (SEQ ID NO: 5362), IEYGHDSPHKS (SEQ ID NO: 5363), ADYGHDSPHKS (SEQ ID NO: 5364), IET GHDSPHKS (SEQ ID NO: 5365), MEWGHDSPHKS (SEQ ID NO: 5366), CEYGHDSPHKS (SEQ ID NO: 5367), RINGHDSPHKS (SEQ ID NO: 5368), MEIGHDSPHKS (SEQ ID NO: 5369), LEYGHDSPHKS (SEQ ID NO: 5370), ADWGHDSPHKS (SEQ ID NO: 5371), IEIGHDSPHKS (SEQ ID NO: 5372), TIKDNDSPHKS (SEQ ID NO: 5373), DIMGHDSPHKS (SEQ ID NO: 5374), FEQGHDSPHKS (SEQ ID NO: 5375), MEFGHDSPH KS (Sequence No. 5376), CDQGHDSPHKS (Sequence No. 5377), LPEGHDSPHKS (Sequence No. 5378), IENGHDSPHKS (Sequence No. 5379), MESGHDSPHKS (Sequence No. 5380), AEIGHDSPHKS (Sequence No. 5381), VEYGHDSPHKS (Sequence No. 5382), TSNGDDSPHKS (Sequence No. 5383), IEVGHDSPHKS (Sequence No. 5384), MEMGHDSPHKS (Sequence No. 5385), AEVGHDSPHKS (Sequence No. 5386), MDAGHDSPHKS (Sequence No. (Sequence No. 5387), VEWGHDSPHKS (Sequence No. 5388), AEQGHDSPHKS (Sequence No. 5389), LEWGHDSPHKS (Sequence No. 5390), MELGHDSPHKS (Sequence No. 5391), METGHDSPHKS (Sequence No. 5392), MEAGHDSPHKS (Sequence No. 5393), TINRQRSPHKS (Sequence No. 5394), IESGHDSPHKS (Sequence No. 5395), TAKDHDSPHKS (Sequence No. 5396), MEVGHDSPHKS (Sequence No. 5397), CEIGHDSPHKS (Sequence No. 5398),ATNGHDSPHKS (SEQ ID NO: 5399), MDGGHDSPHKS (SEQ ID NO: 5400), QEVGHDSPHKS (SEQ ID NO: 5401), ADQGHDSPHKS (SEQ ID NO: 5402), NMNGHDSPHKS (SEQ ID NO: 5403), TPWEHDSPHKS (SEQ ID NO: 5404), IEMGHDSPHKS (SEQ ID NO: 5405), TANEHDSPHKS (SEQ ID NO: 5406), TINGHDSPHKS (SEQ ID NO: 5407), QQQGHDSPHKS (SEQ ID NO: 5408), TPQDHDSPHKS (SEQ ID NO: 5409), HDWGHDSPHKS (SEQ ID NO: 5410), IEGGHDSPHKS (SEQ ID NO: 5411), (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particle including any one of embodiments 147 to 149.

[0190] 151. Any one of the AAV particles of Embodiments 147 to 150, wherein the AAV capsid variant further comprises one, two, three, four, or all of the following amino acids, numbered according to Sequence ID No. 138: non-Q at position 456 (e.g., R or L), non-N at position 457 (e.g., H, K, or R), non-Q at position 458 (e.g., R or T), non-Q at position 459 (H), and / or non-T at position 460 (N or S).

[0191] 152. Any one of the AAV particles from Embodiments 147 to 151, wherein the AAV capsid variant further contains R at position 456, numbered according to Sequence ID No. 138. 153. Any one of the AAV particles from Embodiments 147 to 151, wherein the AAV capsid variant further comprises L at position 456, numbered according to Sequence ID No. 138.

[0192] 154. Any one of the AAV particles from Embodiments 147 to 153, wherein the AAV capsid variant further comprises H at position 457 and R at position 458, numbered according to Sequence ID No. 138.

[0193] 155. Any one of the AAV particles from Embodiments 147 to 153, wherein the AAV capsid variant further comprises K at position 457 and N at position 460, numbered according to Sequence ID No. 138.

[0194] 156. Any one of the AAV particles from Embodiments 147 to 153, wherein the AAV capsid variant further comprises T at position 458, H at position 459, and S at position 460, as numbered according to Sequence ID No. 138.

[0195] 157. Any one of the AAV particles from Embodiments 147 to 151, wherein the AAV capsid variant further comprises R at position 456, R at position 457, and R at position 458, numbered according to Sequence ID No. 138.

[0196] 158. A single AAV particle from any one of embodiments 147 to 157, wherein [A] and [B] are present in loop IV. 159. An AAV particle from any one of embodiments 147 to 158, wherein [A] is located immediately after position 449, as numbered according to sequence number 138.

[0197] 160. Any one of the AAV particles from Embodiments 147 to 159, wherein [A] replaces positions 450 to 453 (e.g., T450, I451, N452, G453) numbered according to Sequence ID No. 138.

[0198] 161. Any one of the AAV particles of Embodiments 147 to 160, wherein [A] is located immediately after position 449, which is numbered according to Sequence ID No. 138, and [A] replaces positions 450 to 453 (e.g., T450, I451, N452, G453).

[0199] 162. Any one of the AAV particles from Embodiments 147 to 161, wherein [A][B] replaces positions 450 to 455 (e.g., T450, I451, N452, G453, S454, G455) numbered according to Sequence ID No. 138.

[0200] 163. Any one of the AAV particles of Embodiments 147 to 162, wherein [A][B] is located immediately after position 449, numbered according to Sequence ID No. 138, and [A][B] replaces positions 450 to 455 (e.g., T450, I451, N452, G453, S454, G455).

[0201] 164. Any one of the AAV particles of Embodiments 147 to 163, wherein [B] is located immediately after [A] and replaces positions 454 and 455 (e.g., S454 and G455) numbered according to Sequence ID No. 138.

[0202] 165. An AAV particle of any one of embodiments 147 to 164, wherein [B] is located immediately after position 455, as numbered according to sequence numbers 4, 36, 981, or 982. 166. An AAV particle from any one of embodiments 147 to 165, wherein [B] is located immediately after [A].

[0203] 167. An AAV particle from any one of embodiments 147 to 166, wherein the AAV capsid variant comprises [A] and [B] from the N-terminus to the C-terminus. 168. Adeno-associated virus (AAV) particles comprising a viral genome containing an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is given by the following formula: [N1]-[N2]-[N3] (SEQ ID NO: 6407) (wherein, (i)[N1] includes X1, X2, and X3, where X2 is S and X3 is G; (ii)[N2] contains the amino acid sequence SPH; and (iii)[N3] includes X4, X5, and X6, where X5 is K. Adeno-associated virus (AAV) particles containing an amino acid sequence having the following characteristics.

[0204] 169. (i) X4 of [N3] is S, T, N, or A; and (ii) X5 of [N3] is A, V, T, S, G, R, L, or N; An AAV particle of Embodiment 168, wherein the AAV capsid variant optionally includes an amino acid modification of either of the aforementioned amino acids in (i) or (ii), e.g., a conservative substitution.

[0205] 170. AAV particle of embodiment 168 or 169, wherein X4 is S and / or X5 is A. 171. An AAV particle of any one of embodiments 168 to 170, wherein [N3] comprises SK, TK, NK, AK, KA, KV, KT, KS, KG, KR, KL, or KN.

[0206] 172. An AAV particle of any one of Embodiments 168 to 171, wherein [N3] is or comprises SKA, SKV, SKT, SKS, SKG, SKR, TKA, NKA, SKL, SKN, or AKA.

[0207] 173. An AAV particle from any one of embodiments 168 to 172, wherein [N3] is or contains an SKA. 174. An AAV particle from any one of embodiments 168 to 173, wherein [N2]-[N3] comprises SPHSK (SEQ ID NO: 4701), SPHTK (SEQ ID NO: 4725), SPHNK (SEQ ID NO: 4726), or SPHAK (SEQ ID NO: 4727).

[0208] 175. [N2]-[N3], (i) SPHSKA (sequence number 941), SPHSKV (sequence number 4737), SPHSKT (sequence number 4731), SPHSKS (sequence number 962), SPHSKG (sequence number 4732), SPHSKR (sequence number 978), SPHTKA (sequence number 4739), SPHNKA (sequence number 4734), SPHSKL (sequence number 960), SPHSKN (sequence number 4735), or SPHAKA (sequence number 4736); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 168 to 174.

[0209] 176. An AAV particle from any one of embodiments 168 to 175, wherein [N2]-[N3] is SPHSKA (SEQ ID NO: 941) or contains the same. 177. Any one of the AAV particles from Embodiments 168 to 176, wherein the AAV capsid variant comprises an amino acid other than G at position 453 (e.g., M, T, I, E, S, A, N, V, L, K, H, P, R, W, or D), numbered according to Sequence ID No. 138 or 981.

[0210] 178. An AAV particle of any one of embodiments 168 to 177, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or 981 and contains amino acid G at position 453.

[0211] 179. X1 of [N1] is G, M, T, I, E, S, A, N, V, L, K, H, P, R, W, or D; optionally, the AAV capsid variant is one of any one of embodiments 168 to 178, comprising an amino acid modification of any of the aforementioned amino acids, e.g., a conservative substitution.

[0212] 180. AAV particles of any one of embodiments 168 to 179, wherein [N1] comprises SG, GS, MS, TS, IS, ES, SS, AS, NS, VS, LS, KS, HS, PS, RS, WS, or DS.

[0213] 181. An AAV particle of any one of Embodiments 168 to 180, wherein [N1] is or includes GSG, MSG, TSG, ISG, ESG, SSG, ASG, NSG, VSG, LSG, KSG, HSG, PSG, RSG, WSG, or DSG.

[0214] 182. An AAV particle from any one of embodiments 168 to 181, wherein [N1] is or contains GSG. 183. An AAV particle from any one of embodiments 168 to 182, wherein [N1]-[N2] contains SGSPH (Sequence ID 4752).

[0215] 184. [N1]-[N2], (i) GSGSPH (sequence number 4695), MSGSPH (sequence number 4798), TSGSPH (sequence number 4800), ISGSPH (sequence number 4801), ESGSPH (sequence number 4803), SSGSPH (sequence number 4804), ASGSPH (sequence number 4806), NSGSPH (sequence number 4807), VSGSPH (sequence number 4786), LSGSPH (sequence number 4808), KSGSPH (sequence number 4810), HSGSPH (sequence number 4811), PSGSPH (sequence number 4813), RSGSPH (sequence number 4815), WSGSPH (sequence number 4817), DSGSPH (sequence number 4818); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 168 to 183.

[0216] 185. [N1]-[N2]-[N3] is (i) GSGSPHSKA (SEQ ID NO: 4697), GSGSPHSKV (SEQ ID NO: 4956), MSGSPHSKA (SEQ ID NO: 4957), TSGSPHSKA (SEQ ID NO: 4959), ISGSPHSKA (SEQ ID NO: 4960), GSGSPHSKT (SEQ ID NO: 4962), ESGSPHSKA (SEQ ID NO: 4963), SSGSPHSKA (SEQ ID NO: 4964), GSGSPHSKS (SEQ ID NO: 4953), ASGSPHSKA (SEQ ID NO: 4966), NSGSPHSKA (SEQ ID NO: 4967), VSGSPHSKA (SEQ ID NO: 4913), LSGSPHSKA (SEQ ID NO: 4968) , KSGSPHSKA (sequence number 4970), GSGSPHSKG (sequence number 4972), GSGSPHSKR (sequence number 4945), HSGSPHSKA (sequence number 4973), PSGSPHSKA (sequence number 4975), RSGSPHSKA (sequence number 4977), GSGSPHTKA (sequence number 4978), WSGSPHSKA (sequence number 4980), DSGSPHSKA (sequence number 4981), GSGSPHNKA (sequence number 4983), GSGSPHSKL (sequence number 4943), GSGSPHSKN (sequence number 4994), or GSGSPHAKA (sequence number 4995); (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 168 to 184.

[0217] 186. An AAV particle according to any one of embodiments 168 to 185, wherein [N1]-[N2]-[N3] is GSGSPHSKA (SEQ ID NO: 4697) or contains the same. 187. An AAV capsid variant according to any one of embodiments 168 to 186, containing an amino acid other than Q (e.g., R, P, H, L, K, I, G, S, M, or E) at position 456, an amino acid other than N (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M) at position 457, an amino acid other than Q (e.g., R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, or D) at position 458, an amino acid other than Q (e.g., H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G) at position 459, and / or an amino acid other than T (e.g., I, N, S, H, R, L, D, Y, A, or Q) at position 460, numbered according to SEQ ID NO: 138.

[0218] 188. An AAV particle according to any one of embodiments 168 to 187, wherein the AAV capsid variant contains an amino acid other than Q (e.g., R, P, H, L, K, I, G, S, M, or E) at position 462, an amino acid other than N (e.g., D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M) at position 463, an amino acid other than Q (e.g., R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, or D) at position 464, an amino acid other than Q (e.g., H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G) at position 465, and / or an amino acid other than T (e.g., I, N, S, H, R, L, D, Y, A, or Q) at position 466, numbered according to SEQ ID NO: 981.

[0219] 189. An AAV particle according to any one of embodiments 168 to 188, wherein the AAV capsid variant contains amino acid Q at position 456, amino acid N at position 457, amino acid Q at position 458, amino acid Q at position 459, and / or amino acid T at position 460, numbered according to SEQ ID NO: 138.

[0220] 190. An AAV particle of any one of embodiments 168 - 189, wherein the AAV capsid variant comprises amino acid Q at position 462, amino acid N at position 463, amino acid Q at position 464, amino acid Q at position 465, and / or amino acid T at position 466, numbered according to SEQ ID NO: 981.

[0221] 191. The AAV capsid variant further comprises [N4], and [N4] comprises X7, X8, X9, X10, and X11, (a) X7 is Q, R, P, H, L, K, I, G, S, M, or E; (b) X8 is N, D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M; (c) X9 is Q, R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, D; (d) X10 is Q, H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G; and (e) X11 is T, I, N, S, H, R, L, D, Y, A, Q; Optionally, an AAV particle of any one of embodiments 168 - 190, wherein the AAV capsid variant comprises an amino acid modification of any of the aforementioned amino acids in (a) - (e), such as a conservative substitution.

[0222] 192. [N4] is (i) QNQQT (SEQ ID NO: 5412), QNRHT (SEQ ID NO: 5413), RDQQT (SEQ ID NO: 5414), PNLQT (SEQ ID NO: 5415), HVRQT (SEQ ID NO: 5416), PNQHT (SEQ ID NO: 5417), QSQQT (SEQ ID NO: 5418), QNQQI (SEQ ID NO: 5419), QPAKT (SEQ ID NO: 5420), QTQQN (SEQ ID NO: 5421), QNLAT (SEQ ID NO: 5422), QNQLT (SEQ ID NO: 5423), QGQQT (SEQ ID NO: 5424), LNRQS (SEQ ID NO: 5425), HNQQT (SEQ ID NO: 5426), QNPPT (SEQ ID NO: 5427), QNLQT (SEQ ID NO: 5428), QYQQT (SEQ ID NO: 5429), QDQET (SEQ ID NO: 5430), QNHQT (SEQ ID NO: 5431), QDQQT (SEQ ID NO: 5432), HWQQT (SEQ ID NO: 5433), PNQQT (SEQ ID NO: 5434), QNQLI (SEQ ID NO: 5435), PEQQT (SEQ ID NO: 5436), QRTMT (SEQ ID NO: 5437), QNQQH (SEQ ID NO: 5438), LHQHT (SEQ ID NO: 5439), QHRIT (SEQ ID NO: 5440), QYIHT (SEQ ID NO: 5441), QKFET (SEQ ID NO: 5442 ), QFPST (SEQ ID NO: 5443), HNQQR (SEQ ID NO: 5444), QAIKT (SEQ ID NO: 5445), QNRQT (SEQ ID NO: 5446), QYQHT (SEQ ID NO: 5447), QNPQS (SEQ ID NO: 5448), QHQLT (SEQ ID NO: 5449), QSPPT (SEQ ID NO: 5450), QAKLT (SEQ ID NO: 5451), KSQQT (SEQ ID NO: 5452), QDRPT (SEQ ID NO: 5453), QSQQL (SEQ ID NO: 5454), QAFHT (SEQ ID NO: 5455), QKQQD (SEQ ID NO: 5456), QNAQT (SEQ ID NO: 5457), HNQLT (SEQ ID NO: 5457) Column number 5458), QNQQY (sequence number 5459), QKLNT (sequence number 5460), QNVQT (sequence number 5461), QAQQT (sequence number 5462), QNLQA (sequence number 5463), QTPPT (sequence number 5464), QYQHA (sequence number 5465), QGQQA (sequence number 5466), QPPAT (sequence number 5467), QERPT (sequence number 5468), QDLQT (sequence number 5469), QAMHT (sequence number 5470), LNQQT (sequence number 5471), QHPST (sequence number 5472), PGLQT (sequence number 5473),QGIRT (SEQ ID NO: 5474), QAPAT (SEQ ID NO: 5475), QSQQI (SEQ ID NO: 5476), QIPPT (SEQ ID NO: 5477), QTQLT (SEQ ID NO: 5478), QAPST (SEQ ID NO: 5479), QNTYA (SEQ ID NO: 5480), QNQHI (SEQ ID NO: 5481), QNHLT (SEQ ID NO: 5482), QIGMT (SEQ ID NO: 5483), LNKQT (SEQ ID NO: 5484), QLQQT (SEQ ID NO: 5485), QRMST (SEQ ID NO: 5486), QGILT (SEQ ID NO: 5487), QDRQT (SEQ ID NO: 5488), RDWQT (SEQ ID NO: 5488) Number 5489), QNTHD (SEQ ID NO: 5490), PNLQI (SEQ ID NO: 5491), QERST (SEQ ID NO: 5492), QNYQT (SEQ ID NO: 5493), QRTCT (SEQ ID NO: 5494), QIGHT (SEQ ID NO: 5495), QGAIT (SEQ ID NO: 5496), QVPPT (SEQ ID NO: 5497), QVQQI (SEQ ID NO: 5498), LMRQT (SEQ ID NO: 5499), QYSVT (SEQ ID NO: 5500), QAITT (SEQ ID NO: 5501), QKTLT (SEQ ID NO: 5502), QNQWT (SEQ ID NO: 5503), QLHHT (SEQ ID NO: 5504), QNIII (SEQ ID NO: 5505), QGHHT (SEQ ID NO: 5506), QSKVT (SEQ ID NO: 5507), QLPST (SEQ ID NO: 5508), IGKQT (SEQ ID NO: 5509), QAIHT (SEQ ID NO: 5510), QHGLT (SEQ ID NO: 5511), QFMCT (SEQ ID NO: 5512), QHLQT (SEQ ID NO: 5513), QNHQN (SEQ ID NO: 5514), QPART (SEQ ID NO: 5515), QSLQT (SEQ ID NO: 5516), QSQLT (SEQ ID NO: 5517), QDRQS (SEQ ID NO: 5518), QMPST (SEQ ID NO: 5519), QGSLT (SEQ ID NO: 5519) Number 5520), QVPAT (sequence number 5521), QDKQT (sequence number 5522), HYQQT (sequence number 5523), QVPST (sequence number 5524), RGEQT (sequence number 5525), PGQQT (sequence number 5526), ​​QSLQI (sequence number 5527), LEQQT (sequence number 5528), QNQST (sequence number 5529), QKVIT (sequence number 5530), QNNDQ (sequence number 5531), QSVHT (sequence number 5532), QPLGT (sequence number 5533), HNQET (sequence number 5534), QNLQI (sequence number 5535),QIQQT (sequence number 5536), QVRNT (sequence number 5537), PSNQT (sequence number 5538), QVGHT (sequence number 5539), QRDIT (sequence number 5540), QMPNT (sequence number 5541), RGLQT ​​(sequence number 5542), QKQQT (sequence number 5543), PSLQT (sequence number 5544), QRDQT (sequence number 5545), QAKGT (sequence number 5546), QSAHT (sequence number 5547), QSTMT (sequence number 5548), QREMT (sequence number 5549), QYRAT (sequence number QQT (sequence number 5550), QWQQT (sequence number 5551), QRMNT (sequence number 5552), GDSQT (sequence number 5553), QKIST (sequence number 5554), PSMQT (sequence number 5555), SPRQT (sequence number 5556), MEQQT (sequence number 5557), QYQNT (sequence number 5558), QHQQT (sequence number 5559), INQQT (sequence number 5560), PNQQH (sequence number 5561), ENRQT (sequence number 5562), QTQQA (sequence number 5563), or QNQAT (sequence number 5564); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, or four amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 191, which are or include the same.

[0223] 193. [N1]-[N2]-[N3]-[N4] is (i) The amino acid sequence of either SEQ ID NO: 200 or 2887-3076; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence containing 1, 2, or 3 modifications but no more than 4 modifications relative to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing 1, 2, or 3 different amino acids but no more than 4 different amino acids relative to any one of the amino acid sequences in (i). An AAV particle of embodiment 191 or 192 that is or contains such an amino acid sequence.

[0224] 194. An AAV particle of any one of embodiments 191 - 193, wherein [N1]-[N2]-[N3]-[N4] is GSGSPHSKAQNQQT (SEQ ID NO: 200) or contains the same.

[0225] 195. An AAV particle of any one of embodiments 191 - 193, wherein [N1]-[N2]-[N3]-[N4] is VSGSPHSKAQNQQT (SEQ ID NO: 903) or contains the same.

[0226] 196. An AAV particle of any one of embodiments 168 - 195, wherein the AAV capsid variant contains, at position 449 (numbered according to SEQ ID NO: 138 or 981), an amino acid other than K (e.g., T, E, or N), at position 450 an amino acid other than T (e.g., S, E, A, N, V, Q, or G), at position 451 an amino acid other than I (e.g., F, E, V, L, D, S, C, T, A, N, H, R, G, or W), and / or at position 452 an amino acid other than N (e.g., I, P, K, R, H, S, M, Q, D, T, L, A, Y, V, F, E, W, or G).

[0227] 197. An AAV particle of any one of embodiments 168 - 196, wherein the AAV capsid variant contains, at position 449 (numbered according to SEQ ID NO: 138 or 981), the amino acid K, at position 450 the amino acid T, at position 451 the amino acid I, and / or at position 452 the amino acid N.

[0228] 198. The AAV capsid variant further contains [N0], and [N0] contains X A X B X C and X D and contains (a)X A However, it is K, T, E, or N; (b)X B However, it is T, S, E, A, N, V, Q, or G; (c)X C is I, F, E, V, L, D, S, C, T, A, N, H, R, G, or W; and (d)X D is N, I, P, K, R, H, S, M, Q, D, T, L, A, Y, V, F, E, W, or G; Optionally, one AAV particle of any of Embodiments 168 to 197, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (d), e.g., a conservative substitution.

[0229] 199. [N0] is, (i) KTII (sequence number 5565), KTFP (sequence number 5566), KTEK (sequence number 5567), KTVN (sequence number 5568), KTFN (sequence number 5569), KTIN (sequence number 5570), TTIN (sequence number 5571), KSIN (sequence number 5572), KTER (sequence number 5573), KELH (sequence number 5574), KAIN (sequence number 5575), KTDN (sequence number 5576), KTFH (sequence number 5577), KTSN (sequence number 5578), ETIN (sequence number 5579), NTIN (sequence number 5580), KTEN ( SEQ ID NO: 5581), KTSS (SEQ ID NO: 5582), KTCN (SEQ ID NO: 5583), KTEH (SEQ ID NO: 5584), KAEM (SEQ ID NO: 5585), KATN (SEQ ID NO: 5586), KAIK (SEQ ID NO: 5587), KTDK (SEQ ID NO: 5588), KTFK (SEQ ID NO: 5589), KSDQ (SEQ ID NO: 5590), KTEI (SEQ ID NO: 5591), KTID (SEQ ID NO: 5592), KNTN (SEQ ID NO: 5593), KTET (SEQ ID NO: 5594), KTEL (SEQ ID NO: 5595), KNIN (SEQ ID NO: 5596), KTEA (SEQ ID NO: 5597) ), KTAN (sequence number 5598), NTIY (sequence number 5599), KTFS (sequence number 5600), KTES (sequence number 5601), KTTN (sequence number 5602), KTED (sequence number 5603), KTNN (sequence number 5604), KEVH (sequence number 5605), KTIS (sequence number 5606), KTVR (sequence number 5607), KTDR (sequence number 5608), ETIK (sequence number 5609), KNHI (sequence number 5610), KESD (sequence number 5611), KTIK (sequence number 5612), KTDL (sequence number 5613), KTVP (distribution Column number 5614), KTVI (sequence number 5615), KAEH (sequence number 5616), KNCL (sequence number 5617), KTVK (sequence number 5618), KNAD (sequence number 5619), KTIT (sequence number 5620), KNCV (sequence number 5621), KNAL (sequence number 5622), KVIN (sequence number 5623), KTEF (sequence number 5624), KTRE (sequence number 5625), KQGE (sequence number 5626), KSEK (sequence number 5627), KNVN (sequence number 5628), KGGE (sequence number 5629), KEFV (sequence number 5630),KSDK (SEQ ID NO: 5631), KTEQ (SEQ ID NO: 5632), KEVQ (SEQ ID NO: 5633), KTEY (SEQ ID NO: 5634), KNCW (SEQ ID NO: 5635), KTDV (SEQ ID NO: 5636), KSDI (SEQ ID NO: 5637), KNSI (SEQ ID NO: 5638), KNSL (SEQ ID NO: 5639), KEVV (SEQ ID NO: 5640), KTEP (SEQ ID NO: 5641), KSEL (SEQ ID NO: 5642), KTWQ (SEQ ID NO: 5643), KTEV (SEQ ID NO: 5644), KAVN (SEQ ID NO: 5631) 5645), KGVL (sequence number 5646), KTEG (sequence number 5647), KTRD (sequence number 5648), KTGN (sequence number 5649), KNAI (sequence number 5650), KAEN (sequence number 5651), KAET (sequence number 5652), KTVH (sequence number 5653), KETA (sequence number 5654), KNNL (sequence number 5655), EAIN (sequence number 5656), KSLN (sequence number 5657), KTIP (sequence number 5658), or KTIH (sequence number 5659); (ii) Any portion of the amino acid sequence in (i), for example, any two or three amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 198, which are or include the same.

[0230] 200. [N0]-[N1]-[N2]-[N3]-[N4] is (i) One amino acid sequence from sequence numbers 3239-3526 or 3591-3605; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence containing 1, 2, or 3 modifications but no more than 4 modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing 1, 2, or 3 different amino acids but no more than 4 different amino acids relative to any one of the amino acid sequences in (i) An AAV particle of Embodiment 198 or 199 that is or contains the above.

[0231] 201. An AAV particle of any one of Embodiments 198 - 200, wherein [N0]-[N1]-[N2]-[N3]-[N4] is KTINGSGSPHSKAQNQQT (SEQ ID NO: 5660) or contains the same.

[0232] [[ID=twelve]]202. An AAV particle of any one of Embodiments 198 - 200, wherein [N0]-[N1]-[N2]-[N3]-[N4] is KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589) or contains the same.

[0233] 203. An adeno - associated virus (AAV) particle containing a viral genome that includes an AAV capsid variant (e.g., an AAV9 capsid variant) and a syntaxin - binding protein 1 (STXBP1) coding sequence (e.g., encoding a human STXBP1 protein), wherein the AAV capsid variant has an amino acid sequence of the following formula: [N1]-[N2]-[N3] (SEQ ID NO: 6408) (wherein, (i) [N1] includes X1, X2, and X3, X2 is an amino acid other than S, and X3 is an amino acid other than G; (ii) [N2] includes the amino acid sequence SPH; and (iii) [N3] includes X4, X5, and X6, and X\(_4\) is K) An adeno - associated virus (AAV) particle containing the above - described amino acid sequence.

[0234] 204. (i) X5 of [N3] is S, I, T, R, H, Y, L, or M; and (ii) X6 of [N3] is G, A, L, E, V, R, W, N, Q, or K; An AAV particle of Embodiment 203, wherein the AAV capsid variant optionally includes an amino acid modification of either of the aforementioned amino acids in (i) or (ii), e.g., a conservative substitution.

[0235] 205. AAV particle of embodiment 203 or 204, wherein X5 is S and / or X6 is G. 206. An AAV particle from any one of embodiments 203 to 205, wherein [N3] comprises KS, KI, KT, KR, KH, KY, KL, KM, SG, IG, TG, RG, SA, SL, SE, SV, SR, SW, SN, HG, YG, SQ, IV, SK, LW, MG, or MA.

[0236] 207. An AAV particle from any one of embodiments 203 to 206, wherein [N3] is or comprises KSG, KIG, KTG, KRG, KSA, KSL, KSE, KSV, KSR, KSW, KSN, KHG, KYG, KSQ, KIV, KSK, KLW, KMG, or KMA.

[0237] 208. An AAV particle of any one of embodiments 203 to 207, wherein [N3] is or contains KSG. 209. Any one of the AAV particles from Embodiments 203 to 208, wherein [N2]-[N3] includes SPHKS (SEQ ID NO: 4704), SPHKI (SEQ ID NO: 4713), SPHKT (SEQ ID NO: 4711), SPHKR (SEQ ID NO: 4717), NPHKS (SEQ ID NO: 5661), SPHKH (SEQ ID NO: 4728), SPHKY (SEQ ID NO: 4715), SPHKL (SEQ ID NO: 4714), or SPHKM (SEQ ID NO: 4729).

[0238] 210. [N2]-[N3] (i) SPHKSG (sequence number 946), SPHKIG (sequence number 958), SPHKTG (sequence number 4738), SPHKRG (sequence number 974), NPHKSG (sequence number 5662), SPHKSA (sequence number 977), SPHKSL (sequence number 4740), SPHKSE (sequence number 4741), SPHKSV (sequence number 4742), SPHKSR (sequence number 951), SPHKSW (sequence number 4743), SPHKSN (sequence number 4744), SPHKHG (sequence number 4745), SPHKYG (sequence number 966), SPHKSQ (sequence number 4746), SPHKIV (sequence number 5663), SPHKSK (sequence number 4747), SPHKLW (sequence number 4748), SPHKMG (sequence number 4750), or SPHKMA (sequence number 4751); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 203 to 209.

[0239] 211. Any one of the AAV particles from Embodiments 203 to 210, wherein [N2]-[N3] is SPHKSG (SEQ ID NO: 946) or contains the same. 212. Any one of the AAV particles from Embodiments 203 to 211, wherein the AAV capsid variant comprises an amino acid other than G at position 453 (e.g., A, K, W, R, L, I, M, N, T, E, Q, Y, H, F, or V), numbered according to Sequence ID No. 138 or 981.

[0240] 213. Any one of the AAV particles from Embodiments 203 to 212, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or 981 and contains amino acid G at position 453.

[0241] 214. (i) X1 in [N1] is G, A, K, W, R, L, I, M, N, T, E, Q, Y, H, F, or V; (ii) X2 in [N1] is H, Y, R, Q, N, P, or D; (iii) X3 of [N1] is D, E, G, V, or N; Optionally, one AAV particle of any of Embodiments 203 to 214, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (i), (ii), or (iii), e.g., a conservative substitution.

[0242] 215. Any one of the AAV particles of Embodiments 203 to 214, wherein X2 of [N1] is H and X3 of [N1] is D. 216. Any one of the AAV particles from embodiments 203 to 215, wherein X1 of [N1] is G, X2 of [N1] is H, and X3 of [N1] is D.

[0243] 217. Any one of the AAV particles from Embodiments 203 to 216, wherein [N1] comprises GH, HD, GY, GR, GQ, AH, GN, KH, GP, WH, RH, LH, IH, MH, GD, NH, TH, EH, QH, YH, HH, FH, VH, YD, HE, RG, QD, RD, ND, PD, QV, DD, HN, or NG.

[0244] 218. Any one of the AAV particles of Embodiments 203 to 217, wherein [N1] is or includes GHD, GYD, GHE, GRG, GQD, GRD, AHD, GND, KHD, GPD, WHD, RHD, LHD, GQV, IHD, MHD, GDD, GHN, NHD, THD, GNG, EHD, QHD, YHD, HHD, FHD, or VHD.

[0245] 219. An AAV particle from any one of embodiments 203 to 218, wherein [N1] is or contains a GHD. 220. An AAV particle from any one of embodiments 203 to 219, wherein [N1]-[N2] contains HDSPH (SEQ ID NO: 4703).

[0246] 221. [N1]-[N2], (i) GHDSPH (SEQ ID NO: 4784), GYDSPH (SEQ ID NO: 4829), GHESPH (SEQ ID NO: 4793), GRGSPH (SEQ ID NO: 4788), GHDNPH (SEQ ID NO: 5664), GQDSPH (SEQ ID NO: 4785), GRDSPH (SEQ ID NO: 4831), AHDSPH (SEQ ID NO: 5665), GNDSPH (SEQ ID NO: 4832), KHDSPH (SEQ ID NO: 5666), GPDSPH (SEQ ID NO: 4833), WHDSPH (SEQ ID NO: 5667), RHDSPH (SEQ ID NO: 5668), LHDSPH (SEQ ID NO: 5669) GQVSPH (sequence number 4835), IHDSPH (sequence number 5670), MHDSPH (sequence number 5671), GDDSPH (sequence number 4792), GHNSPH (sequence number 4836), NHDSPH (sequence number 5672), THDSPH (sequence number 5673), GNGSPH (sequence number 4805), EHDSPH (sequence number 5674), QHDSPH (sequence number 5675), YHDSPH (sequence number 5676), HHDSPH (sequence number 5677), FHDSPH (sequence number 5678), or VHDSPH (sequence number 5679); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, four, or five amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 203 to 220.

[0247] 222. [N1]-[N2]-[N3] is (i) GHDSPHKSG (SEQ ID NO: 4698), GHDSPHKIG (SEQ ID NO: 4996), GYDSPHKSG (SEQ ID NO: 4997), GHESPHKSG (SEQ ID NO: 4998), GHDSPHKTG (SEQ ID NO: 4999), GRGSPHKRG (SEQ ID NO: 5000), GHDNPHKSG (SEQ ID NO: 5680), GQDSPHKSG (SEQ ID NO: 4908), GHDSPHKSA (SEQ ID NO: 4940), GHDSPHKSL (SEQ ID NO: 5001), GHDSPHKSE (SEQ ID NO: 5003), GRDSPHKSG (SEQ ID NO: 5004), AH DSPHKSG (SEQ ID NO: 5681), GNDSPHKSV (SEQ ID NO: 5005), AHDSPHKIG (SEQ ID NO: 5682), GHESPHKSA (SEQ ID NO: 4939), GQDSPHKIG (SEQ ID NO: 5006), GHDSPHKSV (SEQ ID NO: 5007), GHDSPHKSR (SEQ ID NO: 4942), KHDSPHKSG (SEQ ID NO: 5683), GPDSPHKIG (SEQ ID NO: 5008), GPDSPHKSG (SEQ ID NO: 5009), GHDSPHKSW (SEQ ID NO: 5010), WHDSPHKSG (SEQ ID NO: 5684), RHDSPHKS G (SEQ ID NO: 5685), GHDSPHKSN (SEQ ID NO: 5011), GHDSPHKRG (SEQ ID NO: 4937), GHDSPHKHG (SEQ ID NO: 5013), LHDSPHKSG (SEQ ID NO: 5686), GQVSPHKSG (SEQ ID NO: 5014), IHDSPHKSG (SEQ ID NO: 5687), MHDSPHKSG (SEQ ID NO: 5688), GDDSPHKSV (SEQ ID NO: 5015), GHNSPHKSG (SEQ ID NO: 5016), NHDSPHKSG (SEQ ID NO: 5689), THDSPHKSG (SEQ ID NO: 5690), GNGSPHKRG (SEQ ID NO: 5017), EHDSPHKSG (sequence number 5691), GHDSPHKYG (sequence number 5018), GHDSPHKSQ (sequence number 5019), QHDSPHKSG (sequence number 5692), RHDSPHKIV (sequence number 5693), YHDSPHKSG (sequence number 5694), GNDSPHKIG (sequence number 5020), HHDSPHKSG (sequence number 5695), GHDSPHKSK (sequence number 5021), FHDSPHKSG (sequence number 5696), GHDSPHKLW (sequence number 5022), VHDSPHKSG (sequence number 5697),GHDSPHKMG (sequence number 5024), GHDSPHKMA (sequence number 5025), or GDDSPHKSG (sequence number 4938); (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, or 9 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 203 to 221.

[0248] 223. Any one of the AAV particles from Embodiments 203 to 222, wherein [N1]-[N2]-[N3] is GHDSPHKSG (SEQ ID NO: 4698) or contains the same. 224. Any one of the AAV particles from Embodiments 203 to 223, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, comprising an amino acid other than Q at position 456 (e.g., R, P, H, K, L, V, A, E, or I), an amino acid other than N at position 457 (e.g., I, K, S, H, R, T, D, Y, L, W, F, A, Q, or M), an amino acid other than Q at position 458 (e.g., R, V, K, P, Y, H, L, I, E, or M), an amino acid other than Q at position 459 (e.g., H, L, E, P, W, D, I, V, S, K, R, C, M, or N), and / or an amino acid other than T at position 460 (e.g., A, E, K, S, I, P, G, or N).

[0249] 225. Any one of the AAV particles of Embodiments 203 to 224, wherein the AAV capsid variant is numbered according to Sequence ID No. 982, comprising an amino acid other than Q at position 462 (e.g., R, P, H, K, L, V, A, E, or I), an amino acid other than N at position 463 (e.g., I, K, S, H, R, T, D, Y, L, W, F, A, Q, or M), an amino acid other than Q at position 464 (e.g., R, V, K, P, Y, H, L, I, E, or M), an amino acid other than Q at position 465 (e.g., H, L, E, P, W, D, I, V, S, K, R, C, M, or N), and / or an amino acid other than T at position 466 (e.g., A, E, K, S, I, P, G, or N).

[0250] 226. Any one of the AAV particles of Embodiments 203 to 225, wherein the AAV capsid variant comprises amino acid Q at position 456, amino acid N at position 457, amino acid Q at position 458, amino acid Q at position 459, and / or amino acid T at position 460, as numbered according to Sequence ID No. 138.

[0251] 227. Any one of the AAV particles from Embodiments 203 to 226, wherein the AAV capsid variant comprises amino acid Q at position 462, amino acid N at position 463, amino acid Q at position 464, amino acid Q at position 465, and / or amino acid T at position 466, as numbered according to Sequence ID No. 138.

[0252] 228. The AAV capsid variant further comprises [N4], wherein [N4] comprises X7, X8, X9, X10, and X11. (a) X7 is Q, R, P, H, L, K, I, G, S, M, or E; (b) X8 is N, D, V, S, P, T, G, Y, W, E, R, H, K, F, A, I, L, or M; (c) X9 is Q, R, L, A, P, H, T, I, F, K, V, M, G, W, Y, S, E, N, D; (d) X10 is Q, H, K, A, L, P, E, M, I, S, N, R, Y, C, V, T, W, D, G; and (e) X11 is T, I, N, S, H, R, L, D, Y, A, Q; Optionally, one AAV particle of any of Embodiments 203 to 227, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (e), e.g., a conservative substitution.

[0253] 229. [N4] ga, (i) QNQQT (SEQ ID NO: 5412), QIRQT (SEQ ID NO: 5698), QNQHA (SEQ ID NO: 5699), QKQQT (SEQ ID NO: 5543), QSVQT (SEQ ID NO: 5700), RSQQT (SEQ ID NO: 5701), QNKLE (SEQ ID NO: 5702), QNQQK (SEQ ID NO: 5703), QHQQA (SEQ ID NO: 5704), QIQHT (SEQ ID NO: 5705), PRQQT (SEQ ID NO: 5706), HTQQT (SEQ ID NO: 5707), QRQHT (SEQ ID NO: 5708), QSQQT (SEQ ID NO: 5418), QNQQS (SEQ ID NO: 5709), RNQET (SEQ ID NO: 5710), QTQLT (SEQ ID NO: 5478), KNQQT (SEQ ID NO: 5711), QDQQT (SEQ ID NO: 5432), HNQQT (SEQ ID NO: 5426), QNQLT (SEQ ID NO: 5423), QTQQT (SEQ ID NO: 5712), QTQQI (SEQ ID NO: 5713), QSKQA (SEQ ID NO: 5714), QNQPP (SEQ ID NO: 5715), QSPQT (SEQ ID NO: 5716), QNYQT (SEQ ID NO: 5493), QNHQT (SEQ ID NO: 5431), QNRQT (SEQ ID NO: 5446), QNQQG (SEQ ID NO: 5717), QNHLT (SEQ ID NO: 5482) ), QYQHT (SEQ ID NO: 5447), QNQWT (SEQ ID NO: 5503), QNQHT (SEQ ID NO: 5718), QTRQT (SEQ ID NO: 5719), QNLHT (SEQ ID NO: 5720), LNQQT (SEQ ID NO: 5471), QNQET (SEQ ID NO: 5721), QHLQT (SEQ ID NO: 5513), LNQPT (SEQ ID NO: 5722), QNQDT (SEQ ID NO: 5723), RNQQT (SEQ ID NO: 5724), QNLLT (SEQ ID NO: 5725), QLVIT (SEQ ID NO: 5726), RTQET (SEQ ID NO: 5727), QTHQT (SEQ ID NO: 5728), QNQPA (SEQ ID NO: 5728) Column number 5729), QDQHT ​​(sequence number 5730), QSQHT (sequence number 5731), RNQQI (sequence number 5732), VRQQT (sequence number 5733), QNQHS (sequence number 5734), AWQQT (sequence number 5735), QSVPT (sequence number 5736), QNIQP (sequence number 5737), QNHLN (sequence number 5738), LDQQT (sequence number 5739), PDQQS (sequence number 5740), ESQQT (sequence number 5741), QNKQT (sequence number 5742), QRQLT (sequence number 5743), QIIVT (sequence number 5744),QKQST (SEQ ID NO: 5745), QSHQT (SEQ ID NO: 5746), QFVVT (SEQ ID NO: 5747), QNLQT (SEQ ID NO: 5428), QNQQI (SEQ ID NO: 5419), QSQPT (SEQ ID NO: 5748), QNEQT (SEQ ID NO: 5749), QSLQT (SEQ ID NO: 5516), RNRQT (SEQ ID NO: 5750), QSKQT (SEQ ID NO: 5751), QNPLT (SEQ ID NO: 5752), RDQKT (SEQ ID NO: 5753), HNQQN (SEQ ID NO: 5754), QWKRT (SEQ ID NO: 5755), QSQQI (SEQ ID NO: 5476), QAQQT (SEQ ID NO: 5462), QNHQI (SEQ ID NO: 5756), QNQQA (SEQ ID NO: 5757), QNQLN (SEQ ID NO: 5757) (Sequence number 5758), QTQPT (Sequence number 5759), INQQT (Sequence number 5560), QKQLT (Sequence number 5760), RNQLA (Sequence number 5761), RNQQS (Sequence number 5762), ISIQT (Sequence number 5763), QNQQN (Sequence number 5764), QSQQS (Sequence number 5765), QTVCT (Sequence number 5766), QYQQI (Sequence number 5767), QQIMT (Sequence number 5768), QNEQS (Sequence number 5769), LNHQT (Sequence number 5770), QMIHT (Sequence number 5771), RNHQS (Sequence number 5772), QKMNT (Sequence number 5773), QSQQN (Sequence number 5774), QYQHA (Sequence number 5465); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, or four amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 228, which are or include the same.

[0254] 230. [N1]-[N2]-[N3]-[N4] is (i) The amino acid sequence of either SEQ ID NO: 201 or 3160-3237; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 228 or 229, which are or include such particles.

[0255] 231. Any one of the AAV particles of Embodiments 228 to 230, wherein [N1]-[N2]-[N3]-[N4] is or contains GHDSPHKSGQNQQT (Sequence ID 201).

[0256] 232. Any one of the AAV particles from Embodiments 203 to 231, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or 982, comprising an amino acid other than K at position 449 (e.g., T), an amino acid other than T at position 450 (e.g., A, S, I, V, N, E, Y, C, G, W, or Q), an amino acid other than I at position 451 (e.g., E, V, S, T, N, D, C, G, Q, L, P, A), and / or an amino acid other than N at position 452 (e.g., S, Y, I, K, F, T, D, E, G, V, L, A, M, Q, H, P, or R).

[0257] 233. Any one of the AAV particles from Embodiments 203 to 232, wherein the AAV capsid variant comprises amino acid K at position 449, amino acid T at position 450, amino acid I at position 451, and / or amino acid N at position 452, as numbered according to Sequence ID No. 138 or 982.

[0258] 234. The AAV capsid variant further contains [N0], where [N0] is X A , X B , X C , and X D Includes, (a)XA However, it is either K or T; (b)X B However, it is T, A, S, I, V, N, E, Y, C, G, W, or Q; (c)X C However, I, E, V, S, T, N, D, C, G, Q, L, P, A; and (d)X D is N, S, Y, I, K, F, T, D, E, G, V, L, A, M, Q, H, P, or R; Optionally, one AAV particle of any of Embodiments 203 to 233, wherein the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (d), e.g., a conservative substitution.

[0259] 235. [N0] is, (i) KAIN (sequence number 5575), KTIN (sequence number 5570), KTES (sequence number 5601), TTIN (sequence number 5571), KSIN (sequence number 5572), KTVN (sequence number 5568), KSIY (sequence number 5775), KTSN (sequence number 5578), KTTN (sequence number 5602), KIIN (sequence number 5776), KTIS (sequence number 5606), KAII (sequence number 5777), KTIK (sequence number 5612), KTEF (sequence number 5624), KTIT (sequence number 5620), KTNN (sequence number 5604), KTID ( SEQ ID NO: 5592), KAIS (SEQ ID NO: 5778), KTVD (SEQ ID NO: 5779), KTIE (SEQ ID NO: 5780), KTEG (SEQ ID NO: 5647), KVIN (SEQ ID NO: 5623), KAVN (SEQ ID NO: 5645), KTIY (SEQ ID NO: 5781), KTDN (SEQ ID NO: 5576), KTCN (SEQ ID NO: 5583), KNVV (SEQ ID NO: 5782), KTEL (SEQ ID NO: 5595), KTDA (SEQ ID NO: 5783), KTEV (SEQ ID NO: 5644), KSEL (SEQ ID NO: 5642), KTEM (SEQ ID NO: 5784), KTEQ (SEQ ID NO: 5632) ), KTII (sequence number 5565), KIVN (sequence number 5785), KTEK (sequence number 5567), KTEN (sequence number 5581), KIGN (sequence number 5786), KEVM (sequence number 5787), KYQV (sequence number 5788), KTEA (sequence number 5597), KATN (sequence number 5586), KTEH (sequence number 5584), KTVE (sequence number 5789), KAID (sequence number 5790), KTIM (sequence number 5791), KEVG (sequence number 5792), KSEM (sequence number 5793), KAQQ (sequence number 5794), KCGE (distribution Column number 5795), KASN (sequence number 5796), KTET (sequence number 5594), KTIG (sequence number 5797), KTDP (sequence number 5798), KELV (sequence number 5799), KELM (sequence number 5800), KNEI (sequence number 5801), KTPN (sequence number 5802), KITN (sequence number 5803), KTDI (sequence number 5804), KTDQ (sequence number 5805), KGIN (sequence number 5806), KSEI (sequence number 5807), KSEK (sequence number 5627), KWSA (sequence number 5808), KELA (sequence number 5809),KQTQ (SEQ ID NO: 5810), KGAD (SEQ ID NO: 5811), KVGE (SEQ ID NO: 5812), KANE (SEQ ID NO: 5813), KTDT (SEQ ID NO: 5814), KTCI (SEQ ID NO: 5815), KELR (SEQ ID NO: 5816), KCQI (SEQ ID NO: 5817), KGVM (SEQ ID NO: 5818), KACD (SEQ ID NO: 5819), KNEL (SEQ ID NO: 5820), KAAE (SEQ ID NO: 5821), K GQN (sequence code 5822), KNEF (sequence code 5823), KTSI (sequence code 5824), KAEH (sequence code 5616), KCDQ (sequence code 5825), KEIL (sequence code 5826), KTER (sequence code 5573), KNAI (sequence code 5650), KTDK (sequence code 5588), KTPD (sequence code 5827), KTIH (sequence code 5659), or KTEI (sequence code 5591); (ii) Any portion of the amino acid sequence in (i), for example, any two or three amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 234, which are or include such particles.

[0260] 236. [N0]-[N1]-[N2]-[N3]-[N4] is (i) Any one amino acid sequence from sequence numbers 3606 to 3836; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particles of Embodiment 234 or 235, which are or include such particles.

[0261] 237. Any one of the AAV particles from Embodiments 234 to 236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KTINGHDSPHKSGQNQQT (SEQ ID NO: 5828).

[0262] 238. Any one of the AAV particles from Embodiments 234 to 236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KAEIGHDSPHKSGQNQQT (Sequence ID 1754).

[0263] 239. Any one of the AAV particles from Embodiments 234 to 236, wherein [N0]-[N1]-[N2]-[N3]-[N4] is or contains KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241).

[0264] 240. Any one of the AAV particles from Embodiments 168 to 239, wherein [N1]-[N2]-[N3] are present in Loop IV, numbered, for example, according to Sequence ID No. 4, 36, 138, 981, or 982.

[0265] 241. One of the AAV particles from Embodiments 198-202 or 234-240, wherein [N0] and [N4] are present in Loop IV, which is numbered, for example, according to Sequence ID No. 4, 36, 138, 981, or 982.

[0266] 242. One of the AAV particles of Embodiments 198-202 or 234-241, wherein [N0] is located immediately after position 448, according to the amino acid sequence of SEQ ID NOs. 4, 36, 138, 981, or 982.

[0267] 243. One of the AAV particles from Embodiments 198-202 or 234-242, wherein [N0] replaces positions 449-452 (e.g., K449, T450, I451, and N452) numbered according to Sequence ID No. 4, 36, 138, 981, or 982.

[0268] 244. An AAV particle of any one of Embodiments 198-202 or 234-243, wherein [N0] is located immediately after position 448, numbered according to Sequence ID No. 4, 36, 138, 981, or 982, and [N0] replaces positions 449-452 (e.g., K449, T450, I451, and N452).

[0269] 245. An AAV particle from any one of Embodiments 198-202 or 234-244, wherein [N0] corresponds to position 449-452 (e.g., K449, T450, I451, and N452) of any one of Sequence IDs 4, 36, 138, 981, or 982.

[0270] 246. An AAV particle of any one of embodiments 168 to 245, wherein [N1] is located immediately after position 452, as numbered according to sequence numbers 4, 36, 138, 981, or 982.

[0271] 247. An AAV particle of any one of embodiments 168 to 246, wherein [N1] replaces positions 453 to 455 (e.g., G453, S454, and G455) numbered according to sequence numbers 4, 36, 138, 981, or 982.

[0272] 248. An AAV particle of any one of embodiments 168 to 246, wherein [N1] replaces position 453 (e.g., G453) numbered according to sequence numbers 4, 36, 138, 981, or 982.

[0273] 249. (i) X1 in [N1] replaces position 453 (e.g., G453); (ii) X2 of [N1] corresponds to position 454 (for example, S454); and (iii) X3 in [N1] corresponds to the 455th position (for example, G455), (i), (ii), and (iii) are any one of the AAV particles of embodiments 168-177, 179-181, 183-185, 187-193, 195-200, 202, or 240-246, numbered according to sequence number 138 or sequence number 981.

[0274] 250. (i) X1 of [N1] corresponds to position 453 (for example, G453); (ii) X2 of [N1] corresponds to position 454 (for example, S454); and (iii) X3 of [N1] corresponds to position 455 (for example, G455); (i), (ii), and (iii) are any one of the AAV particles of embodiments 168-176, 178-201, or 240-246, numbered according to sequence number 138 or sequence number 981.

[0275] 251. (i) X1 of [N1] corresponds to position 453 (for example, G453); (ii) X2 of [N1] replaces position 454 (e.g., S454); and (iii) X3 of [N1] replaces the 455th position (for example, G455), (i), (ii), and (iii) are any one of the AAV particles of Embodiments 203 to 248, numbered according to Sequence ID No. 138 or Sequence ID No. 982.

[0276] 252. An AAV particle from any one of Embodiments 203-248 or 251, wherein [N1] corresponds to positions 453-455 of Sequence ID No. 982 (e.g., G453, H454, D455).

[0277] 253. An AAV particle from any one of embodiments 168-176, 178-201, 240-247, or 250, where [N1] corresponds to positions 453-455 of sequence number 138 or 981 (e.g., G453, S454, G455).

[0278] 254. An AAV particle of any one of embodiments 168 to 253, wherein [N2] is located immediately after position 455, as numbered according to sequence numbers 4, 36, 138, 981, or 982.

[0279] 255. An AAV particle from any one of embodiments 168 to 254, wherein [N2] corresponds to positions 456 to 458 of sequence number 981 or 982 (e.g., S456, P457, and H458).

[0280] 256. An AAV particle from any one of embodiments 168 to 254, wherein [N2] corresponds to positions 456 to 458 (e.g., S456, P457, and H458) of any one of sequence numbers 4 or 36 to 59.

[0281] 257. Any one of the AAV particles from embodiments 168 to 256, wherein [N2] is located immediately after [N1]. 258. An AAV particle from any one of embodiments 168-202, 240-247, or 249-257, where [N3] corresponds to positions 459-460 of sequence number 981 (e.g., S459, K460, A461).

[0282] 259. One AAV particle from implementations 168-202, 240-247, or 249-257, where [N3] corresponds to the 459-460 position (e.g., S459, K460, A461) of sequence numbers 36, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, or 59.

[0283] 260. An AAV particle of any one of embodiments 168 to 259, wherein [N2]-[N3] is located immediately after position 455, which is numbered according to any one of sequence numbers 4, 36, 138, 981, or 982.

[0284] 261. Any one of the AAV particles from embodiments 168 to 259, wherein [N2]-[N3] is located immediately after position 455, as numbered according to sequence number 981. 262. An AAV particle from any one of embodiments 168 to 261, wherein [N2]-[N3] corresponds to positions 456-461 of sequence number 981 (e.g., S456, P457, H458, S459, K460, A461).

[0285] 263. Any one of the AAV particles from Embodiments 168 to 262, wherein [N2]-[N3] corresponds to position 456-461 (e.g., S456, P457, H458, S459, K460, A461) of any one of sequence numbers 36, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 57, or 59.

[0286] 264. An AAV particle from any one of embodiments 203-257 or 259-261, wherein [N3] corresponds to positions 459-460 of sequence number 982 (e.g., K459, S460, G461).

[0287] 265. An AAV particle from any one of embodiments 203-257 or 259-261, wherein [N3] corresponds to positions 459-460 of sequence number 37 (e.g., K459, S460, G461).

[0288] 266. Any one of the AAV particles from Embodiments 203 to 265, wherein [N2]-[N3] are located immediately after position 455, as numbered according to Sequence ID No. 982. 267. One of the AAV particles of embodiments 203-246, 248, 252, 255, 257, 260, 263-266, in which [N3] replaces positions 454 and 455 (e.g., S454 and G455) numbered according to sequence number 138.

[0289] 268. One of the AAV particles of embodiments 203-246, 248, 252, 255, 257, 260, 263-267, wherein [N3] is located immediately after [N2] and replaces positions 454 and 455 (e.g., S454 and G455) numbered according to sequence number 138.

[0290] 269. One of the AAV particles of Embodiments 203-246, 248, 252, 255, 257, 260, 263-268, wherein [N3] is located immediately after [N1]-[N2] and replaces positions 454 and 455 (e.g., S454 and G455) numbered according to Sequence ID No. 138.

[0291] 270. An AAV particle from any one of embodiments 203-257, 260, 264-269, wherein [N2]-[N3] corresponds to positions 456-461 of sequence number 982 (e.g., S456, P457, H458, K459, S460, G461).

[0292] 271. An AAV particle from any one of embodiments 203-257, 260, or 264-270, wherein [N2]-[N3] corresponds to positions 456-461 of sequence number 37 (e.g., S456, P457, H458, K459, S460, G461).

[0293] 272. One of the AAV particles from Embodiments 191-202 or 228-271, wherein [N4] is located immediately after position 455, as numbered according to Sequence ID No. 138. 273. AAV particle in any one of Embodiments 191-202 or 228-272, wherein [N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460) numbered according to Sequence ID No. 138.

[0294] 274. An AAV particle from any one of Embodiments 191-202 or 228-273, wherein [N4] corresponds to positions 462-466 ​​of Sequence ID No. 981 or 982 (e.g., Q462, N463, Q464, Q465, and T466).

[0295] 275. An AAV particle from any one of Embodiments 191-202 or 228-273, wherein [N4] corresponds to position 462-466 ​​of any one of Sequence ID No. 4 or 36-59. 276. An AAV particle from any one of Embodiments 191-202 or 228-274, wherein [N4] corresponds to positions 456-460 of Sequence ID No. 138 (e.g., Q456, N457, Q458, Q459, and T460).

[0296] 277. AAV particles of any one of Embodiments 191-202 or 228-276, wherein [N2]-[N3]-[N4] replace positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460) numbered according to Sequence ID No. 138.

[0297] 278. An AAV particle in any one of embodiments 191-202 or 228-277, wherein [N2]-[N3]-[N4] is located immediately after position 455, as numbered according to sequence number 138, and [N2]-[N3]-[N4] replaces positions 456-460 (e.g., Q456, N457, Q458, Q459, and T460).

[0298] 279. AAV particles of any one of Embodiments 191-202 or 228-278, wherein [N1]-[N2]-[N3]-[N4] replace positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459, and T460) numbered according to Sequence ID No. 138.

[0299] 280. An AAV particle of any one of Embodiments 191-202 or 228-279, wherein [N1]-[N2]-[N3]-[N4] is located immediately after position 452, which is numbered according to Sequence ID No. 138, and [N1]-[N2]-[N3]-[N4] replaces positions 453-460 (e.g., G453, S454, G455, Q456, N457, Q458, Q459, and T460).

[0300] 281. One of the AAV particles from Embodiments 191-202, 240-247, 249, 250, 253-263, 266, or 272-280, where [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 of Sequence ID No. 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465, and T466).

[0301] 282. One of the AAV particles from Embodiments 168-202, 240-247, 249, 250, 253-263, 266, or 272-280, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 of Sequence ID No. 981 (e.g., G453, S454, G455, S456, P457, H458, S459, K460, A461).

[0302] 283. One of the AAV particles from any of embodiments 228-257, 260, 261, 264-282, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 of sequence number 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465, T466).

[0303] 284. One of the AAV particles from embodiments 203-257, 260, 261, 264-283, wherein [N1]-[N2]-[N3] corresponds to positions 453-461 of sequence number 982 (e.g., G453, H454, D455, S456, P457, H458, K459, S460, G461).

[0304] 285. One of the AAV particles of Embodiments 228-257, 260, 261, 264-282, wherein [N1]-[N2]-[N3]-[N4] corresponds to positions 453-466 of one of Sequence ID No. 4 or 36-59.

[0305] 286. AAV particles of any one of Embodiments 198-202 or 234-286, wherein [N0]-[N1]-[N2]-[N3]-[N4] replace positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460) numbered according to Sequence ID No. 138.

[0306] 287. An AAV particle in any one of Embodiments 198-202 or 234-286, wherein [N0]-[N1]-[N2]-[N3]-[N4] is located immediately after position 448, numbered according to Sequence ID No. 138, and [N0]-[N1]-[N2]-[N3]-[N4] replaces positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and T460).

[0307] 288. One of the AAV particles of Embodiments 198-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, or 287, where [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 of Sequence ID No. 981 (e.g., K449, T450, I451, N452, G453, S454, G455, S456, P457, H458, S459, K460, A461, Q462, N463, Q464, Q465, T466).

[0308] 289. One of the AAV particles of Embodiments 234-257, 260, 261, 264-284, 286, or 287, where [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 of Sequence ID No. 982 (e.g., K449, T450, I451, N452, G453, H454, D455, S456, P457, H458, K459, S460, G461, Q462, N463, Q464, Q465, T466).

[0309] 290. One AAV particle of any one of embodiments 234-257, 260, 261, 264-284, 286, or 287, wherein [N0]-[N1]-[N2]-[N3]-[N4] corresponds to positions 449-466 of any one of sequence numbers 4 or 36-59.

[0310] 291. One of the AAV particles from any one of Embodiments 191-202 or 228-290, wherein [N4] is located immediately after position 461, numbered according to Sequence ID No. 4, 36, 981, or 982.

[0311] 292. One of the AAV particles of Embodiments 191-202 or 228-291, wherein [N4] replaces positions 462-466 ​​(e.g., Q462, N463, Q464, Q465, and T466) numbered according to Sequence ID No. 4, 36, 981, or 982.

[0312] 293. AAV particles in any one of Embodiments 191-202 or 228-292, wherein [N2]-[N3]-[N4] replace positions 462-466 ​​(e.g., Q462, N463, Q464, Q465, and T466) numbered according to Sequence ID No. 4, 36, 981, or 982.

[0313] 294. An AAV particle in any one of Embodiments 191-202 or 228-293, wherein [N2]-[N3]-[N4] is located immediately after position 455, which is numbered according to Sequence ID No. 4, 36, 981, or 982, and [N2]-[N3]-[N4] replaces positions 462-466 ​​(e.g., Q462, N463, Q464, Q465, and T466).

[0314] 295. Any one of the AAV particles from Embodiments 168 to 294, wherein the AAV capsid variant comprises [N2]-[N3] from the N-terminus to the C-terminus. 296. Any one of the AAV particles from Embodiments 168 to 295, wherein the AAV capsid variant comprises [N1]-[N2]-[N3] from the N-terminus to the C-terminus.

[0315] 297. An AAV particle from any one of Embodiments 168 to 296, wherein the AAV capsid variant comprises [N0]-[N1]-[N2]-[N3] from the N-terminus to the C-terminus. 298. An AAV particle from any one of Embodiments 168 to 297, wherein the AAV capsid variant comprises [N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0316] 299. Any one of the AAV particles from Embodiments 168 to 298, wherein the AAV capsid variant comprises [N0]-[N1]-[N2]-[N3]-[N4] from the N-terminus to the C-terminus.

[0317] 300. Adeno-associated virus (AAV) particles comprising a viral genome including an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant comprises formula [A][B] (SEQ ID NO: 4696), (i)[A] contains GSGSPH (sequence number 4695); and (ii)[B] includes X1, X2, X3, X4, and X5, (a) X1 is S, I, F, V, C, Y, W, R, P, L, Q, M, K, or G; (b) X2 is K, M, R, F, V, C, P, Y, L, W, G, N, S, T, I, or A; (c) X3 is A, Y, L, R, W, C, T, F, H, I, P, M, K, S, V, G, Q, or N; (d) X4 is Q, M, F, K, H, R, C, W, P, V, L, G, S, Y, I, A, T, D, N, or E; and (e) X5 is A, N, Y, R, K, L, I, M, Q, S, C, W, F, T, G, V, or P; By choice, adeno-associated virus (AAV) particles in which the AAV capsid variant includes an amino acid modification, e.g., a conservative substitution, of any of the aforementioned amino acids in (a) to (e).

[0318] 301. (a) X1 is S, L, R, V, or P; (b) X2 is K, C, F, L, P, R, S, or V; (c) X3 is A, C, F, I, K, L, M, P, R, T, W, or Y; (d) X4 is Q, R, S, T, C, F, K, L, P or Y; and (e) X5 is N, R, S, T, K, M, Q, or Y; An AAV particle of Embodiment 300 in which, optionally, the AAV capsid variant includes an amino acid modification of any of the aforementioned amino acids in (a) to (e), e.g., a conservative substitution.

[0319] 302. [B]が、SKA、SMY、SKL、SKR、SKW、SRC、SFT、SKF、IVW、SKY、SCH、FPW、SKI、VYY、S LY、SKP、SRF、SRM、SVK、SWA、SLW、SFR、SKK、SYA、SCS、SGA、SFP、SFF、SMC、SKT SGK, FYR, CRV, YGI, VNC, SLA, WSY, RWL, PSC, SSW, SKG, VPW, SGC, STT, PKR, SKC, WVP, SFW, RIK, SKM, LRW, LPT, SYM, LLC, RCC, LCV, SYL, QGC, MAF, SFQ, SLC 、RPW、RPR、SCP、SVR、SLP、VYH、SYT、LVY、YRY、SWL、CPA、SPP、RWT、PRK、PFV、S KS、WVA、SKV、CAL、SSC、SKN、LCT、STC、SKQ、KSG、SYY、SLT、SCQ、FPF、SVF、GRY 、AQA、AQN、YMN、AFY、LKR、RHR、AQK、WRL、CRN、TCN、FFI、AQY、WQN、YFM、ARQ、H QN、IRR、YQN、YWN、AFS、FWN、AQC、MRN、KKN、APN、WKN、ARW、RPN、KVF、AFN、ACS 、RLW、SRN、CPN、ACN、FRQ、PFN、FGN、CQN、LFW、TRK、KRN、RQN、VQN、IQN、AQR、P FR、AWN、RSY、LQN、WLN、RRA、AQT、GCT、RYT、TPN、ARM、CFL、PQN、WSN、FKN、KQN 、APR、RYN、MIC、TQN、WKS、AAR、LTR、IRG、LVN、FQN、ACQ、WGL、ILR、QIN、ACI、A LR、AHA、CLN、AFV、AQF、RCN、MPC、KTS、PYN、AQS、TRN、LKN、AQM、CTN、PDN、RNY ACR, CSV, ARI, LPK, SEQ, VRM, NSR, RKR, ARN, QRP, RVV, GQN, YSN, QSN, AKG, CTS, FEN, AKK, KAQ, MYM, KAF, KLK, KRH, KWR, RCR, FTC, KFF, VWQ, KYF, KAR, CHQ 、PWQ、KIR、YYQ、LYW、KPK、RFW、RMR、VKK、WAP、LWK、FRP、KKV、YAF、KAC、KRL、C SR、RCP、GAC、KFR、FFG、MCQ、KLF、KTR、GKR、YRQ、RVQ、GIQ、NCQ、KPF、LAW、KRS、SYQ、WLQ、KRR、KGC、KRY、GCQ、FTP、TTC、KRQ、KCF、VPQ、FWS、KFK、IKQ、KAP、FRY、KMI、RWQ、KPTQ、YMRY 、KAA、LCQ、CCQ、CVQ、KLT、KLC、YLV、AFQ、KWG、KIL、FQI、KAL、KAH、LCL、PRQ、CPQ、VRY、MPVRC、KLPK、KLT Y, YHQ, YTR, VYQ, RYQ, WLK, PAQ, MCT, PPD, WTQ, RKQ, KCS, FVQ, KLP, KSE, VAQ, LYQ, KVR, SALQ, KNS, KNS RK, CTQ, TCL, YAR, KQR, KRV, SGQ, YYS, LTC, CQS, KAK KPQ, PFQ, KCT, VFE, 300, 301, AAV and 301

[0320] 303. [B] includes SKAQ (sequence number 5829), SMYM (sequence number 5830), SKAF (sequence number 5831), SKLK (sequence number 5832), SKRH (sequence number 5833), SKWR (sequence number 5834), SRCR (sequence number 5835), SFTC (sequence number 5836), SKFF (sequence number 5837), IVWQ (sequence number 5838), SKYF (sequence number 5839), SKAR (sequence number 5840), SCHQ (sequence number 5841), FPWQ (sequence number 5842), SKIR (sequence number 5843), VYYQ (sequence number 5844), SLY W (SEQ ID NO: 5845), SKPK (SEQ ID NO: 5846), SRFW (SEQ ID NO: 5847), SRMR (SEQ ID NO: 5848), SVKK (SEQ ID NO: 5849), SWAP (SEQ ID NO: 5850), SLWK (SEQ ID NO: 5851), SFRP (SEQ ID NO: 5852), SKKV (SEQ ID NO: 5853), SYAF (SEQ ID NO: 5854), SKAC (SEQ ID NO: 5855), SKRL (SEQ ID NO: 5856), SCSR (SEQ ID NO: 5857), SRCP (SEQ ID NO: 5858), SGAC (SEQ ID NO: 5859), SKFR (SEQ ID NO: 5860), SFPF (SEQ ID NO: 586 1) SFFG (SEQ ID NO: 5862), SMCQ (SEQ ID NO: 5863), SKLF (SEQ ID NO: 5864), SKTR (SEQ ID NO: 5865), SGKR (SEQ ID NO: 5866), FYRQ (SEQ ID NO: 5867), CRVQ (SEQ ID NO: 5868), YGIQ (SEQ ID NO: 5869), VNCQ (SEQ ID NO: 5870), SKPF (SEQ ID NO: 5871), SLAW (SEQ ID NO: 5872), SKRS (SEQ ID NO: 5873), WSYQ (SEQ ID NO: 5874), RWLQ (SEQ ID NO: 5875), PSCQ (SEQ ID NO: 5876), SSWL (SEQ ID NO: 5877), SKRR (SEQ ID NO: 5877) Column number 5878), SKGC (sequence number 5879), VPWQ (sequence number 5880), SKRY (sequence number 5881), SGCQ (sequence number 5882), SFTP (sequence number 5883), STTC (sequence number 5884), PKRQ (sequence number 5885), SKCF (sequence number 5886), WVPQ (sequence number 5887), SFWS (sequence number 5888), SKFK (sequence number 5889), RIKQ (sequence number 5890), SKAP (sequence number 5891), SFRY (sequence number 5892), SKMI (sequence number 5893), LRWQ (sequence number 5894),LPTQ (SEQ ID NO: 5895), SKWK (SEQ ID NO: 5896), SYMR (SEQ ID NO: 5897), SKAA (SEQ ID NO: 5898), LLCQ (SEQ ID NO: 5899), RCCQ (SEQ ID NO: 5900), LCVQ (SEQ ID NO: 5901), SKLT (SEQ ID NO: 5902), SKLC (SEQ ID NO: 5903), SYLV (SEQ ID NO: 5904), QGCQ (SEQ ID NO: 5905), MAFQ (SEQ ID NO: 5906), SKWG (SEQ ID NO: 5907), SKIL (SEQ ID NO: 5908), SFQI (SEQ ID NO: 5909), SKAL (SEQ ID NO: 5910), SKAH (SEQ ID NO: 5906) Number 5911), SLCL (sequence number 5912), RPWQ (sequence number 5913), RPRQ (sequence number 5914), SCPQ (sequence number 5915), SVRY (sequence number 5916), SVRC (sequence number 5917), SKMP (sequence number 5918), SKKT (sequence number 5919), SLPY (sequence number 5920), VYHQ (sequence number 5921), SYTR (sequence number 5922), LVYQ (sequence number 5923), YRYQ (sequence number 5924), SWLK (sequence number 5925), CPAQ (sequence number 5926), SMCT (sequence number 5927), SPPD (SEQ ID NO: 5928), SKRN (SEQ ID NO: 5929), RWTQ (SEQ ID NO: 5930), PRKQ (SEQ ID NO: 5931), SKCS (SEQ ID NO: 5932), PFVQ (SEQ ID NO: 5933), SKLP (SEQ ID NO: 5934), SKSE (SEQ ID NO: 5935), WVAQ (SEQ ID NO: 5936), SLYQ (SEQ ID NO: 5937), SKVR (SEQ ID NO: 5938), CALQ (SEQ ID NO: 5939), SSCT (SEQ ID NO: 5940), SKNS (SEQ ID NO: 5941), SKRK (SEQ ID NO: 5942), LCTQ (SEQ ID NO: 5943), STCL (SEQ ID NO: 5943) Number 5944), SYAR (sequence number 5945), SKQR (sequence number 5946), SKRV (sequence number 5947), KSGQ (sequence number 5948), SYYS (sequence number 5949), SLTC (sequence number 5950), SCQS (sequence number 5951), SKAK (sequence number 5952), SKPQ (sequence number 5953), FPFQ (sequence number 5954), SKCT (sequence number 5955), SVFE (sequence number 5956), GRYQ (sequence number 5957), KAQA (sequence number 5958), KAQN (sequence number 5959), MYMN (sequence number 5960),KAFY (sequence number 5961), KLKR (sequence number 5962), KRHR (sequence number 5963), KAQK (sequence number 5964), KWRL (sequence number 5965), RCRN (sequence number 5966), FTCN (sequence number 5967), KFFI (sequence number 5968), KAQY (sequence number 5969), VWQN (sequence number 5970), KYFM (sequence number 5971), KARQ (sequence number 5972), CHQN (sequence number 5973), PWQN (sequence number 5974), KIRR (sequence number 5975), YYQN (sequence number 5976), LYWN (sequence number) Number 5977), KPKR (SEQ ID NO: 5978), KAFS (SEQ ID NO: 5979), RFWN (SEQ ID NO: 5980), KAQC (SEQ ID NO: 5981), RMRN (SEQ ID NO: 5982), VKKN (SEQ ID NO: 5983), WAPN (SEQ ID NO: 5984), LWKN (SEQ ID NO: 5985), KARW (SEQ ID NO: 5986), FRPN (SEQ ID NO: 5987), KKVF (SEQ ID NO: 5988), YAFN (SEQ ID NO: 5989), KACS (SEQ ID NO: 5990), KRLW (SEQ ID NO: 5991), CSRN (SEQ ID NO: 5992), RCPN (SEQ ID NO: 5993), GACN (sequence number 5994), KFRQ (sequence number 5995), FPFN (sequence number 5996), FFGN (sequence number 5997), MCQN (sequence number 5998), KLFW (sequence number 5999), KTRK (sequence number 6000), GKRN (sequence number 6001), YRQN (sequence number 6002), RVQN (sequence number 6003), GIQN (sequence number 6004), KAQR (sequence number 6005), NCQN (sequence number 6006), KPFR (sequence number 6007), LAWN (sequence number 6008), KRSY (sequence number 6009), SYQN (sequence number) Number 6010), WLQN (sequence number 6011), SCQN (sequence number 6012), SWLN (sequence number 6013), KRRA (sequence number 6014), KAQT (sequence number 6015), KGCT (sequence number 6016), KRYT (sequence number 6017), GCQN (sequence number 6018), FTPN (sequence number 6019), TTCN (sequence number 6020), KARM (sequence number 6021), KRQN (sequence number 6022), KCFL (sequence number 6023), VPQN (sequence number 6024), FWSN (sequence number 6025), KFKN (sequence number 6026),IKQN (sequence number 6027), KAPR (sequence number 6028), FRYN (sequence number 6029), KMIC (sequence number 6030), RWQN (sequence number 6031), PTQN (sequence number 6032), KWKS (sequence number 6033), YMRN (sequence number 6034), KAAR (sequence number 6035), LCQN (sequence number 6036), CCQN (sequence number 6037), CVQN (sequence number 6038), KLTR (sequence number 6039), KLCT (sequence number 6040), KIRG (sequence number 6041), YLVN (sequence number 6042), AFQN (sequence number) Number 6043), KACQ (SEQ ID NO: 6044), KWGL (SEQ ID NO: 6045), KILR (SEQ ID NO: 6046), FQIN (SEQ ID NO: 6047), KACI (SEQ ID NO: 6048), KALR (SEQ ID NO: 6049), KAHA (SEQ ID NO: 6050), LCLN (SEQ ID NO: 6051), KAFV (SEQ ID NO: 6052), PRQN (SEQ ID NO: 6053), CPQN (SEQ ID NO: 6054), KAQF (SEQ ID NO: 6055), VRYN (SEQ ID NO: 6056), VRCN (SEQ ID NO: 6057), KMPC (SEQ ID NO: 6058), KKTS (SEQ ID NO: 6059), LPYN (sequence code 6060), YHQN (sequence code 6061), KAQS (sequence code 6062), YTRN (sequence code 6063), VYQN (sequence code 6064), RYQN (sequence code 6065), WLKN (sequence code 6066), KAQM (sequence code 6067), PAQN (sequence code 6068), MCTN (sequence code 6069), PPDN (sequence code 6070), KRNY (sequence code 6071), WTQN (sequence code 6072), KACR (sequence code 6073), RKQN (sequence code 6074), KCSV (sequence code 6075), KARI (array Number 6076), FVQN (Sequence ID 6077), KLPK (Sequence ID 6078), KSEQ (Sequence ID 6079), VAQN (Sequence ID 6080), LYQN (Sequence ID 6081), KVRM (Sequence ID 6082), ALQN (Sequence ID 6083), SCTN (Sequence ID 6084), KNSR (Sequence ID 6085), KRKR (Sequence ID 6086), CTQN (Sequence ID 6087), TCLN (Sequence ID 6088), YARN (Sequence ID 6089), KQRP (Sequence ID 6090), KRVV (Sequence ID 6091), SGQN (Sequence ID 6092),AAV particles from any one of Embodiments 300 to 302, including YYSN (SEQ ID NO: 6093), LTCN (SEQ ID NO: 6094), CQSN (SEQ ID NO: 6095), KAKG (SEQ ID NO: 6096), KPQN (SEQ ID NO: 6097), PFQN (SEQ ID NO: 6098), KCTS (SEQ ID NO: 6099), VFEN (SEQ ID NO: 6100), or KAKK (SEQ ID NO: 6101).

[0321] 304. [B] is, (i) SKAQA (SEQ ID NO: 6102), SKAQN (SEQ ID NO: 6103), SMYMN (SEQ ID NO: 6104), SKAFY (SEQ ID NO: 6105), SKLKR (SEQ ID NO: 6106), SKRHR (SEQ ID NO: 6107), SKAQK (SEQ ID NO: 6108), SKWRL (SEQ ID NO: 6109), SRCRN (SEQ ID NO: 6110), SFTCN (SEQ ID NO: 6111), SKFFI (SEQ ID NO: 6112), SKAQY (SEQ ID NO: 6113), IVWQN (SEQ ID NO: 6114), SKYFM (SEQ ID NO: 6115), SKARQ (SEQ ID NO: 6116), SCHQN (SEQ ID NO: 6117), FPWQN (SEQ ID NO: 6118), SKIRR (SEQ ID NO: 6119), VYYQN (SEQ ID NO: 6120), SLYWN (SEQ ID NO: 6121), SKPKR (SEQ ID NO: 6122), SKAFS (SEQ ID NO: 6123), SRFWN (SEQ ID NO: 6124), SKAQC (SEQ ID NO: 6125), SRMRN (SEQ ID NO: 6126), SVKKN (SEQ ID NO: 6127), SWAPN (SEQ ID NO: 6128), SLWKN (SEQ ID NO: 6129), SKARW (SEQ ID NO: 6130), SFRPN (SEQ ID NO: 6131), SKKVF (SEQ ID NO: 6132) ), SYAFN (sequence number 6133), SKACS (sequence number 6134), SKRLW (sequence number 6135), SCSRN (sequence number 6136), SRCPN (sequence number 6137), SGACN (sequence number 6138), SKFRQ (sequence number 6139), SFPFN (sequence number 6140), SFFGN (sequence number 6141), SMCQN (sequence number 6142), SKLFW (sequence number 6143), SKTRK (sequence number 6144), SGKRN (sequence number 6145), FYRQN (sequence number 6146), CRVQN (sequence number 6147), YGIQN (distribution) Column number 6148), SKAQR (sequence number 6149), VNCQN (sequence number 6150), SKPFR (sequence number 6151), SLAWN (sequence number 6152), SRKRSY (sequence number 6153), WSYQN (sequence number 6154), RWLQN (sequence number 6155), PSCQN (sequence number 6156), SSWLN (sequence number 6157), SKRRA (sequence number 6158), SKAQT (sequence number 6159), SKGCT (sequence number 6160), VPWQN (sequence number 6161), SKRYT (sequence number 6162), SGCQN (sequence number 6163),SFTPN (sequence code 6164), STTCN (sequence code 6165), SKARM (sequence code 6166), PKRQN (sequence code 6167), SKCFL (sequence code 6168), WVPQN (sequence code 6169), SFWSN (sequence code 6170), SKFKN (sequence code 6171), RIKQN (sequence code 6172), SKAPR (sequence code 6173), SFRYN (sequence code 6174), SKMIC (sequence code 6175), LRWQN (sequence code 6176), LPTQN (sequence code 6177), SKWKS (sequence code 6178), SYMRN (sequence code) Number 6179), SKAAR (SEQ ID NO: 6180), LLCQN (SEQ ID NO: 6181), RCCQN (SEQ ID NO: 6182), LCVQN (SEQ ID NO: 6183), SKLTR (SEQ ID NO: 6184), SKLCT (SEQ ID NO: 6185), SKIRG (SEQ ID NO: 6186), SYLVN (SEQ ID NO: 6187), QGCQN (SEQ ID NO: 6188), MAFQN (SEQ ID NO: 6189), SKACQ (SEQ ID NO: 6190), SKWGL (SEQ ID NO: 6191), SKILR (SEQ ID NO: 6192), SFQIN (SEQ ID NO: 6193), SKACI (SEQ ID NO: 6194), SKALR (sequence number 6195), SKAHA (sequence number 6196), SLCLN (sequence number 6197), SKAFV (sequence number 6198), RPWQN (sequence number 6199), RPRQN (sequence number 6200), SCPQN (sequence number 6201), SKAQF (sequence number 6202), SVRYN (sequence number 6203), SVRCN (sequence number 6204), SKMPC (sequence number 6205), SKKTS (sequence number 6206), SLPYN (sequence number 6207), VYHQN (sequence number 6208), SKAQS (sequence number 6209), SYTRN (sequence number) Number 6210), LVYQN (sequence number 6211), YRYQN (sequence number 6212), SWLKN (sequence number 6213), SKAQM (sequence number 6214), CPAQN (sequence number 6215), SMCTN (sequence number 6216), SPPDN (sequence number 6217), SKRNY (sequence number 6218), RWTQN (sequence number 6219), SKACR (sequence number 6220), PRKQN (sequence number 6221), SKCSV (sequence number 6222), SKARI (sequence number 6223), PFVQN (sequence number 6224), SKLPK (sequence number 6225),SKSEQ (SEQ ID NO: 6226), WVAQN (SEQ ID NO: 6227), SLYQN (SEQ ID NO: 6228), SKVRM (SEQ ID NO: 6229), CALQN (SEQ ID NO: 6230), SSCTN (SEQ ID NO: 6231), SKNSR (SEQ ID NO: 6232), SKRKR (SEQ ID NO: 6233), LCTQN (SEQ ID NO: 6234), STCLN (SEQ ID NO: 6235), SYARN (SEQ ID NO: 6236), SKQRP (SEQ ID NO: 6237), S KRVV (sequence code 6238), KSGQN (sequence code 6239), SYYSN (sequence code 6240), SLTCN (sequence code 6241), SCQSN (sequence code 6242), SKAKG (sequence code 6243), SKPQN (sequence code 6244), FPFQN (sequence code 6245), SKCTS (sequence code 6246), SVFEN (sequence code 6247), SKAKK (sequence code 6248), or GRYQN (sequence code 6249); (ii) Any portion of the amino acid sequence in (i), for example, any two, three, or four amino acids therein, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 300 to 303.

[0322] 305. [A]-[B] is, (i) GSGSPHSKAQA (SEQ ID NO: 6250), GSGSPHSKAQN (SEQ ID NO: 6251), GSGSPHSMYMN (SEQ ID NO: 6252), GSGSPHSKAFY (SEQ ID NO: 6253), GSGSPHSKLKR (SEQ ID NO: 6254), GSGSPHSKRHR (SEQ ID NO: 6255), GSGSPHSKAQK (SEQ ID NO: 6256), GSGSPHSKWRL (SEQ ID NO: 6257), GSGSPHSRCRN (SEQ ID NO: 6258), GSGSPHSFTCN (SEQ ID NO: 6259), GSGSPHSKFFI (SEQ ID NO: 6260), GSG SPHSKAQY (SEQ ID NO: 6261), GSGSPHIVWQN (SEQ ID NO: 6262), GSGSPHSKYFM (SEQ ID NO: 6263), GSGSPHSKARQ (SEQ ID NO: 6264), GSGSPHSCHQN (SEQ ID NO: 6265), GSGSPHFPWQN (SEQ ID NO: 6266), GSGSPHSKIRR (SEQ ID NO: 6267), GSGSPHVYYQN (SEQ ID NO: 6268), GSGSPHSLYWN (SEQ ID NO: 6269), GSGSPHSKPKR (SEQ ID NO: 6270), GSGSPHSKAFS (SEQ ID NO: 6271), GSGSPHSRF WN (Sequence ID 6272), GSGSPHSKAQC (Sequence ID 6273), GSGSPHSRMRN (Sequence ID 6274), GSGSPHSVKKN (Sequence ID 6275), GSGSPHSWAPN (Sequence ID 6276), GSGSPHSLWKN (Sequence ID 6277), GSGSPHSKARW (Sequence ID 6278), GSGSPHSFRPN (Sequence ID 6279), GSGSPHSKKVF (Sequence ID 6280), GSGSPHSYAFN (Sequence ID 6281), GSGSPHSKACS (Sequence ID 6282), GSGSPHSKRLW (Sequence ID number (Sequence number 6283), GSGSPHSCSRN (Sequence number 6284), GSGSPHSRCPN (Sequence number 6285), GSGSPHSGACN (Sequence number 6286), GSGSPHSKFRQ (Sequence number 6287), GSGSPHSFPFN (Sequence number 6288), GSGSPHSFFGN (Sequence number 6289), GSGSPHSMCQN (Sequence number 6290), GSGSPHSKLFW (Sequence number 6291), GSGSPHSKTRK (Sequence number 6292), GSGSPHSGKRN (Sequence number 6293), GSGSPHFYRQN (Sequence number 6294),GSGSPHCRVQN (SEQ ID NO: 6295), GSGSPHYGIQN (SEQ ID NO: 6296), GSGSPHSKAQR (SEQ ID NO: 6297), GSGSPHVNCQN (SEQ ID NO: 6298), GSGSPHSKPFR (SEQ ID NO: 6299), GSGSPHSLAWN (SEQ ID NO: 6300), GSGSPHSKRSY (SEQ ID NO: 6301), GSGSPHWSYQN (SEQ ID NO: 6302), GSGSPHRWLQN (SEQ ID NO: 6303), GSGSPHPSCQN (SEQ ID NO: 6304), GSGSPHSSWLN (SEQ ID NO: 6305), GSGSP HSKRRA (SEQ ID NO: 6306), GSGSPHSKAQT (SEQ ID NO: 6307), GSGSPHSKGCT (SEQ ID NO: 6308), GSGSPHVPWQN (SEQ ID NO: 6309), GSGSPHSKRYT (SEQ ID NO: 6310), GSGSPHSGCQN (SEQ ID NO: 6311), GSGSPHSFTPN (SEQ ID NO: 6312), GSGSPHSTTCN (SEQ ID NO: 6313), GSGSPHSKARM (SEQ ID NO: 6314), GSGSPHPKRQN (SEQ ID NO: 6315), GSGSPHSKCFL (SEQ ID NO: 6316), GSGSPHWVPQN (SEQ ID NO: 6317), GSGSPHSFWSN (SEQ ID NO: 6318), GSGSPHSKFKN (SEQ ID NO: 6319), GSGSPHRIKQN (SEQ ID NO: 6320), GSGSPHSKAPR (SEQ ID NO: 6321), GSGSPHSFRYN (SEQ ID NO: 6322), GSGSPHSKMIC (SEQ ID NO: 6323), GSGSPHLRWQN (SEQ ID NO: 6324), GSGSPHLPTQN (SEQ ID NO: 6325), GSGSPHSKWKS (SEQ ID NO: 6326), GSGSPHSYMRN (SEQ ID NO: 6327), GSGSPHSKAAR (SEQ ID NO: 6327) 6328), GSGSPHLLCQN (sequence number 6329), GSGSPHRCCQN (sequence number 6330), GSGSPHLCVQN (sequence number 6331), GSGSPHSKLTR (sequence number 6332), GSGSPHSKLCT (sequence number 6333), GSGSPHSKIRG (sequence number 6334), GSGSPHSYLVN (sequence number 6335), GSGSPHQGCQN (sequence number 6336), GSGSPHMAFQN (sequence number 6337), GSGSPHSKACQ (sequence number 6338), GSGSPHSKWGL (sequence number 6339),GSGSPHSKILR (SEQ ID NO: 6340), GSGSPHSFQIN (SEQ ID NO: 6341), GSGSPHSKACI (SEQ ID NO: 6342), GSGSPHSKALR (SEQ ID NO: 6343), GSGSPHSKAHA (SEQ ID NO: 6344), GSGSPHSLCLN (SEQ ID NO: 6345), GSGSPHSKAFV (SEQ ID NO: 6346), GSGSPHRPWQN (SEQ ID NO: 6347), GSGSPHRPRQN (SEQ ID NO: 6348), GSGSPHSCPQN (SEQ ID NO: 6349), GSGSPHSKAQF (SEQ ID NO: 6350), GSGSP HSVRYN (SEQ ID NO: 6351), GSGSPHSVRCN (SEQ ID NO: 6352), GSGSPHSKMPC (SEQ ID NO: 6353), GSGSPHSKKTS (SEQ ID NO: 6354), GSGSPHSLPYN (SEQ ID NO: 6355), GSGSPHVYHQN (SEQ ID NO: 6356), GSGSPHSKAQS (SEQ ID NO: 6357), GSGSPHSYTRN (SEQ ID NO: 6358), GSGSPHLVYQN (SEQ ID NO: 6359), GSGSPHYRYQN (SEQ ID NO: 6360), GSGSPHSWLKN (SEQ ID NO: 6361), GSGSPHSKAQM (SEQ ID NO: 6362), GSGSPHCPAQN (SEQ ID NO: 6363), GSGSPHSMCTN (SEQ ID NO: 6364), GSGSPHSPPDN (SEQ ID NO: 6365), GSGSPHSKRNY (SEQ ID NO: 6366), GSGSPHRWTQN (SEQ ID NO: 6367), GSGSPHSKACR (SEQ ID NO: 6368), GSGSPHPRKQN (SEQ ID NO: 6369), GSGSPHSKCSV (SEQ ID NO: 6370), GSGSPHSKARI (SEQ ID NO: 6371), GSGSPHPFVQN (SEQ ID NO: 6372), GSGSPHSKLPK (SEQ ID NO: 6362) 6373), GSGSPHSKSEQ (sequence number 6374), GSGSPHWVAQN (sequence number 6375), GSGSPHSLYQN (sequence number 6376), GSGSPHSKVRM (sequence number 6377), GSGSPHCALQN (sequence number 6378), GSGSPHSSCTN (sequence number 6379), GSGSPHSKNSR (sequence number 6380), GSGSPHSKRKR (sequence number 6381), GSGSPHLCTQN (sequence number 6382), GSGSPHSTCLN (sequence number 6383), GSGSPHSYARN (sequence number 6384),GSGSPHSKQRP (sequence number 6385), GSGSPHSKRVV (sequence number 6386), GSGSPHKSGQN (sequence number 6387), GSGSPHSYYSN (sequence number 6388), GSGSPHSLTCN (sequence number 6389), GSGSPHSCQSN (sequence number 6390), GSGSPHSKAKG (sequence number 6391), GSGSPHSKPQN (sequence number 6392), GSGSPHFPFQN (sequence number 6393), GSGSPHSKCTS (sequence number 6394), GSGSPHSVFEN (sequence number 6395), GSGSPHSKAKK (sequence number 6396), or GSGSPHGRYQN (sequence number 6397); (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) An AAV particle which is or includes one of the embodiments 300 to 304.

[0323] 306. An AAV particle of any one of embodiments 300 to 305, wherein [A]-[B] does not contain the amino acid sequence of GSGSPHSKAQN (SEQ ID NO: 6251). 307. Any one AAV particle of any of Embodiments 300 to 306, wherein the AAV capsid variant comprises one, two, or all of the following, numbered according to Sequence ID No. 138: an amino acid other than Q at position 458 (e.g., R, C, S, W, L, F, Y, H, I, V, A, or P), an amino acid other than Q at position 459 (e.g., K, I, R, L, or S), and / or an amino acid other than T at position 460 (e.g., R).

[0324] 308. AAV capsid variant, (i) Amino acid R at position 458; (ii) Amino acid W at position 458; (iii) Amino acid Y at position 458; (iv) Amino acid F at position 458; (v) Amino acid S at position 458; (vi) Amino acid C at position 458; (vii) Amino acid I at position 458; (viii) Amino acid L at position 458; (ix) Amino acid P at rank 458; (x) Amino acid I at position 459; (xi) Amino acid H at position 458; or (xii) Amino acid V at position 458 Includes; (i) to (xii) are numbered according to sequence number 138. An AAV particle according to any one of embodiments 300 to 307.

[0325] 309. AAV capsid variant, (i) Amino acid R at position 458 and amino acid K at position 459; (ii) Amino acid C at position 458 and amino acid I at position 459; (iii) Amino acid S at position 458 and amino acid R at position 459; (iv) Amino acid L at position 458 and amino acid K at position 459; (v) Amino acid F at position 458 and amino acid K at position 459; (vi) Amino acid C at position 458 and amino acid R at position 459; (vii) Amino acid H at position 458 and amino acid R at position 459; (viii) Amino acid I at position 458 and amino acid L at position 459; (ix) Amino acid V at position 458 and amino acid R at position 459; (x) Amino acid A at position 458 and amino acid K at position 459; (xi) Amino acid I at position 458 and amino acid K at position 459; (xii) Amino acid C at position 458 and amino acid S at position 459; or (xiii) Amino acid C at position 458 and amino acid L at position 459 Includes; (i) to (xiii) are numbered according to sequence number 138. An AAV particle according to any one of embodiments 300 to 307.

[0326] 310. An AAV particle of any one of embodiments 300 to 307, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, comprising amino acid F at position 458, amino acid K at position 459, and amino acid R at position 460.

[0327] 311. Any one AAV particle of Embodiments 300 to 310, wherein the AAV capsid variant comprises one, two, or all of the following, numbered according to Sequence ID No. 138: an amino acid other than T at position 450 (e.g., Y, P, W, R, K, S, or F), an amino acid other than I at position 451 (e.g., R, S, Y, L, V, H, P, A, or F), and / or an amino acid other than N at position 452 (e.g., V, W, A, T, F, Y, L, R, H, S, or M).

[0328] 312. An AAV particle of any one of embodiments 300 to 311, wherein the AAV capsid variant contains amino acid V at position 452, numbered according to Sequence ID No. 138. 313. An AAV particle of any one of Embodiments 300 to 312, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, and comprises amino acid Y at position 450 and amino acid V at position 452.

[0329] 314. An AAV particle of any one of Embodiments 300 to 312, wherein the AAV capsid variant is numbered according to Sequence ID No. 138, and comprises amino acid R at position 450 and amino acid Y at position 451.

[0330] 315. AAV capsid variant, (i) Amino acid P at position 450, amino acid R at position 451, and amino acid W at position 452; (ii) Amino acid Y at position 450, amino acid S at position 451, and amino acid A at position 452; (iii) amino acid Y at position 450, amino acid Y at position 451, and amino acid T at position 452; (iv) Amino acid P at position 450, amino acid R at position 451, and amino acid F at position 452; (v) Amino acid W at position 450, amino acid L at position 451, and amino acid T at position 452; (vi) Amino acid R at position 450, amino acid S at position 451, and amino acid Y at position 452; (vii) Amino acid Y at position 450, amino acid V at position 451, and amino acid F at position 452; (viii) Amino acid K at position 450, amino acid H at position 451, and amino acid L at position 452; (ix) Amino acid P at position 450, amino acid P at position 451, and amino acid L at position 452; (x) Amino acid P at position 450, amino acid A at position 451, and amino acid R at position 452; (xi) Amino acid S at position 450, amino acid R at position 451, and amino acid R at position 452; (xii) Amino acid F at position 450, amino acid F at position 451, and amino acid H at position 452; (xiii) Amino acid R at position 450, amino acid F at position 451, and amino acid S at position 452; (xiv) Amino acid Y at position 450, amino acid S at position 451, and amino acid M at position 452; or (xv) Amino acid P at position 450, amino acid F at position 451, and amino acid L at position 452. Includes; (i) to (xv) are numbered according to sequence number 138. An AAV particle according to any one of embodiments 300 to 311.

[0331] 316. AAV capsid variants, (i) One amino acid sequence from sequence numbers 3849-3982, 2984-4010, or 4681-4693; (ii) Any portion of the amino acid sequence in (i), for example, any 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 amino acids, for example, an amino acid sequence containing consecutive amino acids; (iii) An amino acid sequence that includes one, two, or three but no more than four modifications to any of the amino acid sequences in (i); or (iv) An amino acid sequence containing one, two, or three but no more than four different amino acids for any one of the amino acid sequences in (i) AAV particle comprising any one of embodiments 300 to 315.

[0332] 317. Any one of the AAV particles from Embodiments 300 to 316, wherein the AAV capsid variant does not contain the amino acid sequence GSGSPHSKAQNQQ (SEQ ID NO: 1801) or GSGSPHSKAQNQQT (SEQ ID NO: 200).

[0333] 318. A single AAV particle from any of embodiments 300 to 317, wherein [A]-[B] is present in loop IV. 319. An AAV particle of any one of embodiments 300 to 318, wherein [A] is located immediately after position 452, as numbered according to sequence number 138 or 981.

[0334] 320. An AAV particle of any one of Embodiments 300 to 319, wherein [A] replaces positions 453 to 455 (e.g., G453, S454, G455) numbered according to Sequence ID No. 138 or 981.

[0335] 321. Any one of the AAV particles of Embodiments 300 to 320, wherein [A] is located immediately after position 452, numbered according to Sequence ID No. 138 or 981, and [A] replaces positions 453 to 455 (e.g., G453, S454, G455).

[0336] 322. An AAV particle from any one of embodiments 300 to 321, wherein [B] is located immediately after [A]. 323. An AAV particle of any one of Embodiments 300 to 322, wherein [B] replaces positions 456 and 457 (e.g., Q456, N457) numbered according to Sequence ID No. 138.

[0337] 324. Any one of the AAV particles of Embodiments 300 to 323, wherein [A]-[B] replaces positions 453-457 (e.g., G453, S454, G455, Q456, N457) numbered according to Sequence ID No. 138.

[0338] 325. Any one of the AAV particles of Embodiments 300 to 324, wherein [A]-[B] is located immediately after position 452, numbered according to Sequence ID No. 138, and [A]-[B] replaces positions 453-457 (e.g., G453, S454, G455, Q456, N457).

[0339] 326. An AAV particle of any one of embodiments 300 to 325, wherein the AAV capsid variant comprises [A] and [B] from the N-terminus to the C-terminus. 327. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant comprises at least one, at least two, at least three, or at least four (e.g., from 1 to 4 to 1 to 5) charged amino acid residues (e.g., acidic and / or basic amino acid residues) relative to SEQ ID NO: 138, which are located on the N-terminal side of the amino acid sequence of SPH (e.g., within 1, 2, 3, 4, 5, or 6 amino acids from the start of the SPH amino acid sequence (e.g., within positions 450 to 455 as numbered according to SEQ ID NO: 138)), and optionally, the amino acid sequence of SPH is located at positions 456 to 458 as numbered according to any one of SEQ ID NOs: 36 to 59, 981, or 982.

[0340] 328. An AAV particle of Embodiment 327, wherein the amino acid sequence of SPH is located at positions 456-458, numbered according to one of sequence numbers 36-59, 981, or 982.

[0341] 329. AAV particles of Embodiment 327 or 328, wherein the AAV capsid variant contains fewer than 4, 3, or 2 (e.g., 2 or 1) charged amino acid residues (e.g., acidic and / or basic amino acid residues) relative to SEQ ID NO: 138.

[0342] 330. Any one of the AAV particles of Embodiments 327 to 329, wherein the AAV capsid variant optionally contains one charged amino acid residue (e.g., an acidic or basic amino acid residue) at any one of the positions 450 to 455 numbered relative to Sequence ID No. 138.

[0343] 331. An AAV particle of any one of embodiments 327 to 330, wherein the charged amino acid residue is an acidic amino acid (e.g., D or E). 332. An AAV particle according to any one of embodiments 327 to 331, wherein the charged amino acid residue is a negatively charged amino acid (e.g., D or E).

[0344] 333. An AAV particle of any one of embodiments 327 to 332, wherein the charged amino acid residue is D. 334. An AAV particle of any one of embodiments 327 to 333, wherein the charged amino acid residue is E.

[0345] 335. An AAV particle according to any one of embodiments 327 to 334, wherein the charged amino acid residue is a basic amino acid (e.g., K, R, or H). 336. An AAV particle of any one of embodiments 327 to 335, wherein the charged amino acid residue is a positively charged amino acid (e.g., K, R, or H).

[0346] 337. An AAV particle according to any one of embodiments 327 to 336, wherein the charged amino acid residue is H. 338. An AAV particle of any one of embodiments 327 to 337, wherein the charged amino acid residue is R.

[0347] 339. An AAV particle according to any one of embodiments 327 to 338, wherein the charged amino acid residue is K. 340. An AAV particle of any one of embodiments 327 to 339, wherein the AAV capsid variant comprises an acidic amino acid (e.g., E or D) and a basic amino acid (e.g., R, K, or H).

[0348] 341. Any one of the AAV particles of Embodiments 327 to 340, wherein at least one, two, three, or four charged amino acid residues are located within 1, 2, 3, 4, 5, or 6 (e.g., 1 to 6) amino acids from the beginning of the SPH amino acid sequence.

[0349] 342. Any one of the AAV particles from Embodiments 327 to 341, wherein the AAV capsid variant comprises two charged amino acid residues immediately preceding the SPH amino acid sequence (for example, at positions 454 and 455, numbered according to SEQ ID NO: 138 or SEQ ID NO: 982).

[0350] 343. Any one of the AAV particles from Embodiments 327 to 342, wherein the AAV capsid variant contains a charged amino acid residue (e.g., E) within 1, 2, 3, 4, or 5 (e.g., 5) amino acids from the start of the SPH amino acid sequence.

[0351] 344. An AAV particle of any one of Embodiments 327 to 343, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., E) at position 451, numbered according to any one of Sequence ID No. 138, 981, or 982.

[0352] 345. An AAV particle of any one of embodiments 327 to 344, wherein the AAV capsid variant contains E at position 451, numbered according to any one of sequence numbers 138, 981, or 982.

[0353] 346. An AAV particle of any one of embodiments 327 to 345, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., R or K) at position 452, numbered according to any one of SEQ ID NOs. 138, 981, or 982.

[0354] 347. Any one of the AAV particles from Embodiments 327 to 346, wherein the AAV capsid variant contains R at position 452, numbered according to Sequence ID No. 138 or Sequence ID No. 982. 348. Any one of the AAV particles from Embodiments 327 to 347, wherein the AAV capsid variant is numbered according to Sequence ID No. 138 or Sequence ID No. 982, and comprises E at position 451 and R at position 452.

[0355] 349. Any one of the AAV particles from Embodiments 327 to 348, wherein the AAV capsid variant has reduced directivity to liver cells or tissues compared to the directivity of an AAV capsid containing the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981.

[0356] 350. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant comprises at least one, at least two, at least three, or at least four (e.g., from 1 to 4 to 1 to 5) charged amino acid residues (e.g., basic amino acid residues) relative to SEQ ID NO: 138, which are located on the C-terminal side of the amino acid sequence of SPH (e.g., within 1, 2, 3, 4, 5, 6, or 7 amino acids from the end of the SPH amino acid sequence (e.g., within positions 459 to 465 as numbered according to any one of SEQ ID NOs: 36 to 59 or 981)), and optionally, the amino acid sequence of SPH is located at positions 456 to 458 as numbered according to any one of SEQ ID NOs: 36 to 59, 981, or 982.

[0357] 351. An AAV particle of Embodiment 350, wherein the amino acid sequence of SPH is located at positions 456-458, numbered according to one of sequence numbers 36-59, 981, or 982.

[0358] 352. AAV particles of Embodiment 350 or 351, wherein the AAV capsid variant contains fewer than 4, 3, or 2 (e.g., 2 or 1) charged amino acid residues (e.g., basic amino acid residues) relative to SEQ ID NO: 138.

[0359] 353. Any one of the AAV particles of Embodiments 350 to 352, wherein the AAV capsid variant optionally contains one charged amino acid residue (e.g., a basic amino acid residue) at any one of positions 456 to 460 numbered according to Sequence ID No. 138 or at any one of positions 462 to 466 numbered according to Sequence ID No. 36 to 59, 981, or 982.

[0360] 354. An AAV particle according to any one of embodiments 350 to 353, wherein the charged amino acid residue is a basic amino acid (e.g., R or K). 355. An AAV particle of any one of embodiments 350 to 354, wherein the charged amino acid residue is a positively charged amino acid (e.g., R or K).

[0361] 356. An AAV particle of any one of embodiments 350 to 355, wherein the charged amino acid residue is R. 357. An AAV particle of any one of embodiments 350 to 355, wherein the charged amino acid residue is K.

[0362] 358. Any one of the AAV particles of Embodiments 350 to 357, wherein at least 1, 2, 3, or 4 charged amino acid residues are located within 1, 2, 3, 4, 5, 6, or 7 (e.g., 1 to 7) amino acids from the end of the SPH amino acid sequence.

[0363] 359. Any one of the AAV particles of Embodiments 350 to 358, wherein the AAV capsid variant includes a charged amino acid residue (e.g., K or R) immediately following the SPH sequence (e.g., at position 459, as numbered according to SEQ ID NO: 981).

[0364] 360. Any one of the AAV particles of Embodiments 350 to 359, wherein the AAV capsid variant comprises a charged amino acid residue (e.g., K or R) at position 459, numbered according to SEQ ID NO: 138 or SEQ ID NO: 982.

[0365] 361. Any one of the AAV particles from Embodiments 350 to 360, wherein the AAV capsid variant contains K at position 459, numbered according to Sequence ID No. 981. 362. Any one of the AAV particles of Embodiments 350 to 360, wherein the AAV capsid variant contains R at position 459, numbered according to Sequence ID No. 981.

[0366] 363. Any one of the AAV particles of Embodiments 350 to 362, wherein the AAV capsid variant comprises one, two, three, four, five, or all charged amino acid residues (e.g., R or K) at positions 460, 461, 462, 463, 464, and / or 465, numbered according to Sequence ID No. 138 or 981.

[0367] 364. Any one of the AAV particles from Embodiments 300-326 or 350-363, wherein the AAV capsid variant has reduced directivity to liver cells or tissues compared to the directivity of an AAV capsid containing the amino acid sequence of SEQ ID NO: 138.

[0368] 365. Any one of the AAV particles from Embodiments 300-326 or 350-364, where the AAV capsid variant is concentrated at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90, at least 95, at least 100, at least 105, at least 110, at least 115, at least 120, at least 125, at least 130, at least 135, at least 140, at least 150, at least 160, at least 170, at least 180, at least 190, or at least 200 times in the liver compared to the AAV capsid containing the amino acid sequence of SEQ ID NO: 138, when measured by assay, for example, as described in Example 4.

[0369] 366. Any one of the AAV particles of Embodiments 300-326, 364, or 365, wherein the AAV capsid variant has reduced directivity to CNS cells or tissues, such as brain cells, brain tissue, spinal cord cells, or spinal cord tissue, compared to the directivity of an AAV capsid containing the amino acid sequence of SEQ ID NO: 138.

[0370] 367. Any one of the AAV particles from Embodiments 300-326 or 364-366, wherein the AAV capsid variant exhibits preferential transduction in the liver region compared to transduction in the brain and / or dorsal root ganglia (DRG).

[0371] 368. Any one of the AAV particles of Embodiments 300-326 or 364-367, wherein the AAV capsid variant exhibits preferential transduction in the hepatic region compared to transduction in the heart and / or muscle (e.g., quadriceps).

[0372] 369. Adeno-associated virus (AAV) particles comprising a viral genome containing an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is (a) Any amino acid sequence of the sequences provided in Tables 1, 2A, 2B, or 20-26; (b) an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, or at least 17 consecutive amino acids from any one of the sequences provided in Tables 1, 2A, 2B, or 20-26; or (c) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids for any one of the sequences provided in Tables 1, 2A, 2B, or 20-26; or (d) An amino acid sequence to which any one of the amino acid sequences provided in Tables 1, 2A, 2B, or 20-26 has at least one, at least two, or at least three, but no more than four modifications. Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

[0373] 370. Adeno-associated virus (AAV) particles comprising a viral genome including an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is (a) Any amino acid sequence of sequence numbers 945-980 or 985-986; (b) an amino acid sequence containing at least 3, at least 4, or at least 5 consecutive amino acids from any one of sequence numbers 945-980 or 985-986; or (c) Amino acid sequences containing at least one, at least two, or at least three but no more than four different amino acids for any one amino acid sequence of sequence numbers 945-980 or 985-986; (d) Amino acid sequences that include at least one, at least two, or at least three, but no more than four, modifications to any one of the amino acid sequences of sequence numbers 945-980 or 985-986. Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

[0374] 371. Adeno-associated virus (AAV) particles comprising a viral genome including an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is (a) Any amino acid sequence of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909; (b) an amino acid sequence containing at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive amino acids from any one of sequence numbers 2, 200, 201, 941, 943, 204, 208, 404, or 903-909; (c) For any one amino acid sequence of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909, an amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids; or (d) Amino acid sequences that include at least one, at least two, or at least three, but no more than four, modifications to any one of the amino acid sequences of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909. Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

[0375] 372. Adeno-associated virus (AAV) particles comprising a viral genome including an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is (a) Any amino acid sequence of sequence numbers 3849-4051 or 4681-4693; (b) an amino acid sequence containing at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, or at least 17 consecutive amino acids from any one of sequence numbers 3849-4051 or 4681-4693; (c) an amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids for any one amino acid sequence of sequence numbers 3849-4051 or 4681-4693; or (d) Amino acid sequences that include at least one, at least two, or at least three, but no more than four, modifications to any one of the amino acid sequences of sequence numbers 3849-4051 or 4681-4693 Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

[0376] 373. Adeno-associated virus (AAV) particles comprising a viral genome including an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is (a) Any amino acid sequence from sequence numbers 4052 to 4092; (b) an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, or at least 17 consecutive amino acids from any one of sequence numbers 4052 to 4092; (c) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids for any one amino acid sequence of sequence numbers 4052 to 4092; or (d) Amino acid sequences that include at least one, at least two, or at least three, but no more than four, modifications to any one of the amino acid sequences of sequence numbers 4052 to 4092 Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

[0377] 374. Adeno-associated virus (AAV) particles comprising a viral genome including an AAV capsid variant (e.g., AAV9 capsid variant) and a syntaxin-binding protein 1 (STXBP1) coding sequence (e.g., encoding human STXBP1 protein), wherein the AAV capsid variant is (a) Any amino acid sequence of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097; (b) an amino acid sequence comprising at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16 or at least 17 consecutive amino acids from any one of sequence numbers 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097; (c) For any one amino acid sequence of sequence numbers 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097, an amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids; or (d) Amino acid sequences for any one of the amino acid sequences of SEQ ID NOs: 4056, 4058, 4059, 4062-4064, 4066, 4067, 4080, 4084, 4090, or 4095-4097, which include at least one, at least two, or at least three, but no more than four, modifications. Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

[0378] 375. AAV particles of Embodiment 369 or 371, wherein the AAV capsid variant comprises an amino acid sequence containing at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or at least 13 consecutive amino acids from any one of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.

[0379] 376. An AAV particle from any one of embodiments 369 to 374, comprising at least three consecutive amino acids, including SPH. 377. An AAV particle of any one of embodiments 369-371 or 376, comprising at least four consecutive amino acids, which constitute SPHS (SEQ ID NO: 4700).

[0380] 378. An AAV particle of any one of embodiments 369-371, 376, or 377, comprising at least five consecutive amino acids, including SPHSK (SEQ ID NO: 4701). 379. An AAV particle of any one of embodiments 369-371 or 376-378, comprising at least six consecutive amino acids, including SPHSKA (SEQ ID NO: 941).

[0381] 380. An AAV particle from any one of embodiments 369 to 371, comprising at least three consecutive amino acids, the HDS. 381. An AAV particle of any one of embodiments 369-371 or 380, comprising at least four consecutive amino acids, including HDSP (SEQ ID NO: 4702).

[0382] 382. An AAV particle of any one of embodiments 369-371, 380, or 381, comprising at least five consecutive amino acids, including HDSPH (SEQ ID NO: 4703). 383. An AAV particle comprising at least six consecutive amino acids, comprising HDSPHK (SEQ ID NO: 2), one of the embodiments 369-371 or 380-382.

[0383] 384. (i) At least three consecutive amino acids containing SPH; (ii) At least four consecutive amino acids comprising SPHK (SEQ ID NO: 6398); (iii) at least five consecutive amino acids comprising SPHKY (SEQ ID NO: 4715); and / or (iv) At least six consecutive amino acids containing SPHKYG (SEQ ID NO: 966), An AAV particle from any one of embodiments 369 to 371.

[0384] 385. AAV particles of Embodiment 369 or 371, wherein the AAV capsid variant comprises an amino acid sequence with at least one, at least two, or at least three, but no more than four, modifications to any one of the amino acid sequences of SEQ ID NOs. 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.

[0385] 386. An AAV particle of any one embodiment 369, 371, or 385, wherein the AAV capsid variant comprises an amino acid sequence with at least one, at least two, or at least three, but no more than four, modifications to the amino acid sequence of SPHSKA (SEQ ID NO: 941).

[0386] 387. An AAV particle of any one embodiment 369, 371, or 385, wherein the AAV capsid variant comprises an amino acid sequence with at least one, at least two, or at least three, but no more than four, modifications to the amino acid sequence of HDSPHK (SEQ ID NO: 2).

[0387] 388. An AAV particle of any one of embodiments 369-371, 384, or 385, wherein the AAV capsid variant comprises an amino acid sequence with at least one, at least two, or at least three, but no more than four, modifications to the amino acid sequence of SPHKYG (SEQ ID NO: 966).

[0388] 389. AAV capsid variant, (i) an amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589) that includes at least one, at least two, or at least three but no more than four modifications; (ii) An amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754) that includes at least one, at least two, or at least three, but no more than four, modifications to the amino acid sequence. (iii) An amino acid sequence of KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241) that includes at least one, at least two, or at least three, but no more than four, modifications to the amino acid sequence; (iv) an amino acid sequence of KTINGHDSPHSKAQNLQT (SEQ ID NO: 4100) that includes at least one, at least two, or at least three, but no more than four, modifications; or (v) Amino acid sequence of KTVNGHDSPHSKAQNQQT (SEQ ID NO: 4062) that includes at least one, at least two, or at least three, but no more than four, modifications to the amino acid sequence AAV particles of embodiment 370, including the AAV particles of embodiment 370.

[0389] 390. AAV particles of Embodiment 369 or 371, wherein the AAV capsid variant contains an amino acid sequence that includes at least one, at least two, or at least three, but no more than four, different amino acids for any one of the amino acid sequences of SEQ ID NOs. 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.

[0390] 391. An AAV particle of any one of Embodiments 369, 371, or 390, wherein the AAV capsid variant comprises an amino acid sequence containing at least one, at least two, or at least three, but no more than four, different amino acids relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941).

[0391] 392. An AAV particle of any one embodiment 369, 371, or 390, wherein the AAV capsid variant comprises an amino acid sequence containing at least one, at least two, or at least three, but no more than four, different amino acids relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2).

[0392] 393. An AAV particle from any one of Embodiments 369, 371, 384, or 390, wherein the AAV capsid variant comprises an amino acid sequence containing at least one, at least two, or at least three, but no more than four, different amino acids relative to the amino acid sequence of SPHKYG (SEQ ID NO: 966).

[0393] 394. AAV capsid variant, (i) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids relative to the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589); (ii) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids relative to the amino acid sequence of KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754); (iii) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids relative to the amino acid sequence of KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241); (iv) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids relative to the amino acid sequence of KTINGHDSPHSKAQNLQT (SEQ ID NO: 4100); or (v) An amino acid sequence containing at least one, at least two, or at least three but no more than four different amino acids relative to the amino acid sequence of KTVNGHDSPHSKAQNQQT (SEQ ID NO: 4062) AAV particles of embodiment 369, including the AAV particles of embodiment 369.

[0394] 395. An AAV particle of any one of Embodiments 1-129, 269, 271, 375-388, or 390-394, wherein the AAV capsid variant comprises any one amino acid sequence of SEQ ID NOs: 2, 200, 201, 941, 943, 204, 208, 404, or 903-909.

[0395] 396. One of the AAV particles of Embodiments 295-297, 301-305, 313, 314, 318, 319, or 323, wherein the AAV capsid variant comprises the amino acid sequence ERVSGSPHSKA (SEQ ID NO: 6399), and optionally, the amino acid sequence is located immediately after position 450, numbered according to SEQ ID NO: 138, and replaces positions 451-455 (e.g., I451, N542, G453, S454, G455).

[0396] 397. One of the AAV particles of Embodiments 369-371, 375-379, 385, 386, 389-391, or 394-396, wherein the AAV capsid variant comprises the amino acid sequence KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589), and optionally, the amino acid sequence is located immediately after position 448, numbered according to SEQ ID NO: 138, and replaces positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460).

[0397] 398. One of the AAV particles of Embodiments 269-371, 375, 380-383, 385, 387, 389, 390, 393, or 394, wherein the AAV capsid variant comprises the amino acid sequence AEIGHDSPHKSG (SEQ ID NO: 6400), and optionally, the amino acid sequence is located immediately after position 449, numbered according to SEQ ID NO: 138, and replaces positions 450-455 (e.g., T450, I451, N452, G453, S454, G455).

[0398] 399. An AAV particle of any one of embodiments 369-371, 375, 380-383, 385, 387, 389, 390, 393, 394, or 398, wherein the AAV capsid variant comprises the amino acid sequence KAEIGHDSPHKSGQNQQT (SEQ ID NO: 1754), and optionally, the amino acid sequence is located immediately after position 448 and replaces positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460), numbered according to SEQ ID NO: 138.

[0399] 400. One of the AAV particles of Embodiments 369-371, 375-379, 390, 391, or 395, wherein the AAV capsid variant comprises the amino acid sequence EKMSGSPHSKA (SEQ ID NO: 6401), and optionally, the amino acid sequence is located immediately after position 450, numbered according to SEQ ID NO: 138, and replaces positions 451-455 (e.g., I451, N452, G453, S454, G455).

[0400] 401. One of the AAV particles of Embodiments 369-371, 375-379, 390, 391, or 395, wherein the AAV capsid variant comprises the amino acid sequence KTEKMSGSPHSKAQNQQT (SEQ ID NO: 3241), and optionally, the amino acid sequence is located immediately after position 448, numbered according to SEQ ID NO: 138, and replaces positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460).

[0401] 402. An AAV particle of any one of Embodiments 369-371, 375-379, 390, 391, or 395, wherein the AAV capsid variant comprises the amino acid sequence HDSPHSKAQNL (SEQ ID NO: 6402), and optionally the amino acid sequence is located immediately after position 453, numbered according to SEQ ID NO: 138, and replaces positions 456-458 (e.g., Q456, N457, Q458).

[0402] 403. One of the AAV particles of Embodiments 369-371, 375-379, 390, 391, or 395, wherein the AAV capsid variant comprises the amino acid sequence KTINGHDSPHSKAQNLQT (SEQ ID NO: 4100), and optionally, the amino acid sequence is located immediately after position 448, numbered according to SEQ ID NO: 138, and replaces positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460).

[0403] 404. One of the AAV particles of Embodiments 369-371, 375-379, 390, 391, or 395, wherein the AAV capsid variant comprises the amino acid sequence VNGHDSPHSKA (SEQ ID NO: 6403), and optionally, the amino acid sequence is located immediately after position 450, numbered according to SEQ ID NO: 138, and replaces positions 451-455 (e.g., I451, N452, G453, S454, G455).

[0404] 405. One of the AAV particles of Embodiments 369-371, 375-379, 390, 391, or 395, wherein the AAV capsid variant comprises the amino acid sequence KTVNGHDSPHSKAQNQQT (SEQ ID NO: 4062), and optionally, the amino acid sequence is located immediately after position 448, numbered according to SEQ ID NO: 138, and replaces positions 449-460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, T460).

[0405] 406. The AAV capsid variant is the nucleotide sequence of SEQ ID NO: 942; a nucleotide sequence having at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, e.g., substitutions (e.g., conservative substitutions) to the nucleotide sequence of SEQ ID NO: 942, but with no more than 10 modifications, e.g., substitutions (e.g., conservative substitutions); or at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications to the nucleotide sequence of SEQ ID NO: 942 AAV particle comprising any one of Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, or 395, comprising an amino acid sequence encoded by a nucleotide sequence containing 10 or fewer different nucleotides.

[0406] 407. The AAV capsid variant is the nucleotide sequence of SEQ ID NO: 3; a nucleotide sequence containing at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, e.g., substitutions (e.g., conservative substitutions), but no more than 10 modifications, e.g., substitutions (e.g., conservative substitutions), relative to the nucleotide sequence of SEQ ID NO: 3; or at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, but no more than 10, relative to the nucleotide sequence of SEQ ID NO: 3. AAV particle comprising an amino acid sequence encoded by a nucleotide sequence containing different nucleotides, one of the AAV particles of Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, or 395.

[0407] 408. A nucleotide sequence encoding a capsid variant is the nucleotide sequence of SEQ ID NO: 942; a nucleotide sequence having at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, e.g., substitutions (e.g., conservative substitutions) relative to the nucleotide sequence of SEQ ID NO: 942, but containing 10 or fewer modifications, e.g., substitutions (e.g., conservative substitutions); or having at least one, at least two, at least three, at least four, at least five, at least AAV particles of any one of embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, or 406, comprising a nucleotide sequence containing 6 or at least 7 but 10 or fewer different nucleotides.

[0408] 409. A nucleotide sequence encoding a capsid variant is the nucleotide sequence of SEQ ID NO: 3; a nucleotide sequence having at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications, e.g., substitutions (e.g., conservative substitutions) relative to the nucleotide sequence of SEQ ID NO: 3, but containing no more than 10 modifications, e.g., substitutions (e.g., conservative substitutions); or having at least one, at least two, at least three, at least four, at least five, at least six, or at least seven modifications relative to the nucleotide sequence of SEQ ID NO: 3. AAV particle comprising any one of Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, or 407, comprising a nucleotide sequence containing 10 or fewer different nucleotides.

[0409] 410. Any one of the AAV particles from Embodiments 369 to 409, wherein the amino acid sequence is present in Loop IV, for example, compared to the amino acid sequence of Sequence ID No. 138. 411. Any one of the AAV particles of Embodiments 369 to 410, wherein the amino acid sequence is located immediately after positions 448, 449, 450, 451, 452, 453, 454, or 455, as numbered according to Sequence ID No. 138.

[0410] 412. Any one of the AAV particles of Embodiments 369 to 411, wherein the amino acid sequence replaces amino acids 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, and / or 460 (e.g., K449, T450, I451, N452, G453, S454, G455, Q456, N457, Q458, Q459, and / or T460) numbered according to Sequence ID No. 138.

[0411] 413. Any one of the AAV particles from Embodiments 369 to 412, wherein the amino acid sequence is located immediately after position 455, as numbered according to Sequence ID No. 138. 414. Any one of the AAV particles from Embodiments 369 to 413, wherein the amino acid sequence is located immediately after position 453, as numbered according to Sequence ID No. 138.

[0412] 415. The AAV capsid variant contains the amino acid sequence of SPHSKA (SEQ ID NO: 941), and the amino acid sequence is located immediately after position 455 according to SEQ ID NO: 138, in embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 11 One of the following AAV particles: 5-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, or 410-413.

[0413] 416. The AAV capsid variant contains the amino acid sequence of SPHSKA (SEQ ID NO: 941), and the amino acid sequence is located immediately after position 455, numbered according to SEQ ID NO: 981, in embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 1 One of the following AAV particles: 15-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, or 415.

[0414] 417. AAV particles of Embodiment 415 or 416, wherein the AAV capsid variant further comprises an amino acid other than I at position 451, an amino acid other than N at position 452, and an amino acid other than G at position 453, numbered according to any one of Sequence ID No. 36, 138, or 981.

[0415] 418. Any one of the AAV particles of Embodiments 415 to 417, wherein the AAV capsid variant further comprises E at position 451, R at position 452, and V at position 453, numbered according to any one of Sequence ID No. 36, 138, or 981.

[0416] 419. Any one of the AAV particles of Embodiments 415 to 418, wherein the AAV capsid variant further includes substitutions I451E, N452R, and G453V, which are numbered according to any one of Sequence ID No. 36, 138, or 981.

[0417] 420. AAV capsid variant, (i) E at position 451, R at position 452, and V at position 453, numbered according to one of sequence numbers 36, 138, or 981; and (ii) The amino acid sequence of SPHSKA (SEQ ID NO: 941) (The amino acid sequence is located immediately after position 455, which is numbered according to one of SEQ ID NOs: 36, 138, or 981) AAV particle including any one of embodiments 415 to 419.

[0418] 421. AAV particles of Embodiment 415 or 416, wherein the AAV capsid variant further comprises an amino acid other than I at position 451, an amino acid other than N at position 452, and / or G at position 453, as numbered according to Sequence ID No. 39 or 138.

[0419] 422. Any one of the AAV particles of Embodiment 415, 416, or 421, wherein the AAV capsid variant further comprises E at position 451, K at position 452, and / or M at position 453, as numbered according to Sequence ID No. 138 or 39.

[0420] 423. Any one of the AAV particles of Embodiments 415, 416, 421, or 422, wherein the AAV capsid variant further includes substitutions I451E, N452K, and G453M, which are numbered according to Sequence ID No. 39 or 138.

[0421] 424. AAV capsid variant, (i) Numbered according to sequence number 39 or 138, with E at position 451, K at position 452, and M at position 453; and (ii) The amino acid sequence of SPHSKA (SEQ ID NO: 941) (The amino acid sequence is located immediately after position 455, which is numbered according to SEQ ID NO: 39 or 138) AAV particles including any one of embodiments 415, 416, or 421-423.

[0422] 425. AAV particles of Embodiment 415 or 416, wherein the AAV capsid variant further comprises an amino acid other than S at position 454, an amino acid other than G at position 455, and / or Q at position 458, as numbered according to Sequence ID No. 138.

[0423] 426. Any one of the AAV particles of Embodiment 415, 416, or 425, wherein the AAV capsid variant further comprises H at position 454, D at position 455, and / or L at position 458, as numbered according to Sequence ID No. 138.

[0424] 427. Any one of the AAV particles of Embodiments 415, 416, 425, or 426, wherein the AAV capsid variant further includes substitutions S454H, G455D, and Q458L, numbered according to Sequence ID No. 138.

[0425] 428. AAV capsid variant, (i) H at position 454, D at position 455, and / or L at position 458, as assigned according to sequence number 138; and (ii) The amino acid sequence of SPHSKA (sequence number 941) (the amino acid sequence is located immediately after position 455, which is numbered according to sequence number 138) AAV particles including any one of embodiments 415, 416, or 425-427.

[0426] 429. AAV particles of Embodiment 415 or 416, wherein the AAV capsid variant further comprises an amino acid other than I at position 451, an amino acid other than S at position 454, and / or an amino acid other than G at position 455, as numbered according to Sequence ID No. 52 or 138.

[0427] 430. An AAV particle of any one of Embodiments 415, 416, or 429, wherein the AAV capsid variant further comprises V at position 451, H at position 454, and / or D at position 455, as numbered according to Sequence ID No. 52 or 138.

[0428] 431. Any one of the AAV particles of Embodiments 415, 416, 429, or 430, wherein the AAV capsid variant further includes substitutions I451V, S454H, and / or G455D, which are numbered according to Sequence ID No. 52 or 138.

[0429] 432. AAV capsid variant, (i) V at position 451, H at position 454, and / or D at position 455, numbered according to sequence number 52 or 138; and (ii) The amino acid sequence of SPHSKA (SEQ ID NO: 941) (The amino acid sequence is located immediately after position 455, which is numbered according to SEQ ID NO: 52 or 138) AAV particles including any one of embodiments 415, 416, or 429-431.

[0430] 433. The AAV capsid variant contains the amino acid sequence of HDSPHK (SEQ ID NO: 2), and the amino acid sequence is located immediately after position 453, which is numbered according to the amino acid sequence of SEQ ID NO: 138, in embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-10. Any one of the following AAV particles: 3, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, or 414.

[0431] 434. The AAV capsid variant contains the amino acid sequence of HDSPHK (SEQ ID NO: 2), and the amino acid sequence is located immediately after position 453, which is numbered according to the amino acid sequence of SEQ ID NO: 982, in embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103. Any one of the following AAV particles: 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, or 433.

[0432] 435. The AAV capsid variant contains the amino acid sequence of SPHKSG (SEQ ID NO: 946), and the amino acid sequence is located immediately after position 455, which is numbered according to the amino acid sequence of SEQ ID NO: 982, in embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 1 Any one of the following AAV particles: 05, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433, or 434.

[0433] 436. AAV capsid variant, (i) The amino acid sequence of HDSPHSKA (sequence number 4486) located immediately after position 453; and (ii) Deletion of amino acid SG at positions 454 and 455 Includes; (i) and (ii) are numbered according to sequence number 138. An AAV particle from any one of embodiments 369 to 435.

[0434] 437. Any one of the AAV particles from Embodiments 369 to 436, wherein the AAV capsid variant further comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), numbered according to SEQ ID NO: 138, containing amino acid HD at positions 454 and 455, and present immediately after position 455.

[0435] 438. Any one of the AAV particles of Embodiments 433 to 435, wherein the AAV capsid variant further comprises an amino acid other than T at position 450, an amino acid other than I at position 451, and an amino acid other than N at position 452, as numbered according to Sequence ID No. 138 or 982.

[0436] 439. Any one of the AAV particles of Embodiments 433-435 or 438, wherein the AAV capsid variant further comprises A at position 450, E at position 451, and I at position 452, numbered according to Sequence ID No. 138 or 982.

[0437] 440. Any one of the AAV particles of Embodiments 433-435, 438, or 439, wherein the AAV capsid variant further includes substitutions T450A, I451E, and N452I, which are numbered according to Sequence ID No. 138 or 982.

[0438] 441. AAV capsid variant, (i) A at position 450, E at position 451, and I at position 452, as numbered according to sequence number 138 or 982; and (ii) The amino acid sequence of HDSPHK (SEQ ID NO: 2) (located immediately after position 453, numbered according to SEQ ID NO: 138 or 982) AAV particles including any one of embodiments 433, 434, or 438-440.

[0439] 442. One AAV particle from any of Embodiments 1-22, 25-27, 31, 34-42, 45-50, 53-63, 69, 79, 83-86, 91-98, 102, 103, 110, 111, 118-129, 369-371, 384, 385, 390, 393, 395, 410-413, wherein the AAV capsid variant contains the amino acid sequence of SPHKYG (SEQ ID NO: 966), and the amino acid sequence is located immediately after position 455, which is numbered according to the amino acid sequence of SEQ ID NO: 138.

[0440] 443. Adeno-associated virus (AAV) particles comprising an AAV capsid variant containing the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is located immediately after position 453, numbered according to the amino acid sequence of SEQ ID NO: 982, and the AAV particles further comprising nucleic acids encoding the syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein).

[0441] 444. Adeno-associated virus (AAV) particles comprising nucleic acids encoding an AAV capsid variant and a syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein), wherein the AAV capsid variant comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941), and the amino acid sequence is located immediately after position 455, as numbered according to the amino acid sequence of SEQ ID NO: 981.

[0442] 445. Adeno-associated virus (AAV) particles containing an AAV capsid variant containing the amino acid sequence of HDSPHK (SEQ ID NO: 2), wherein the amino acid sequence is located immediately after position 453, numbered according to the amino acid sequence of SEQ ID NO: 37, and is optionally selected. (i) One, two, or all of the following amino acids, numbered according to Sequence ID No. 138 or 37, that are not T at position 450, not I at position 541, and / or not N at position 452; (ii) One, two, or all of the following, numbered according to sequence number 138 or 37: A at position 450, E at position 451, and / or I at position 452. Including; AAV particles further contain nucleic acids encoding syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein). Adeno-associated virus (AAV) particles.

[0443] 446. Adeno-associated virus (AAV) particles comprising an AAV capsid variant containing the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is located immediately after position 455, numbered according to any one of the amino acid sequences of SEQ ID NOs: 36, 38-55, 57, or 59, and the AAV particles further comprising nucleic acids encoding the syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein).

[0444] 447. AAV capsid variants further, (i) Modifications in Loops I, II, VI and / or VIII; and / or (ii) Substitution at position K449, numbered according to sequence number 138, e.g., K449R substitution AAV particles including any one of the prior embodiments.

[0445] 448. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant comprises an amino acid sequence having at least one, at least two, or at least three modifications, e.g., substitutions (e.g., conservative substitutions) to the amino acid sequence of SEQ ID NO: 138, but with no more than 30, no more than 20, or no more than 10 modifications, e.g., substitutions (e.g., conservative substitutions).

[0446] 449. An AAV particle of any one of the prior embodiments, wherein the AAV capsid variant includes an amino acid sequence containing at least one, at least two, or at least three different amino acids relative to the amino acid sequence of SEQ ID NO: 138, but 30 or fewer, 20 or fewer, or 10 or fewer.

[0447] 450. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant comprises an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with SEQ ID NO: 138.

[0448] 451. An AAV particle of any one of the prior embodiments, wherein the AAV capsid variant comprises an amino acid sequence having at least 98% identity with SEQ ID NO: 138. 452. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant comprises an amino acid sequence encoded by a sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with SEQ ID NO: 137.

[0449] 453. An AAV particle of any one of the preceding embodiments, wherein the nucleotide sequence encoding the capsid variant includes a sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with SEQ ID NO: 137.

[0450] 454. An AAV particle of any one of the preceding embodiments, wherein the AAV capsid variant comprises a VP1 protein, a VP2 protein, a VP3 protein, or a combination thereof.

[0451] 455. Any one of the AAV particles from Embodiments 1 to 454, wherein the AAV capsid variant includes an amino acid sequence corresponding to positions 138-742 of SEQ ID NO: 981, 982, 36, or 4, for example, VP2, or a sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto.

[0452] 456. Any one of the AAV particles from Embodiments 1 to 455, wherein the AAV capsid variant includes an amino acid sequence corresponding to positions 203-742 of SEQ ID NO: 981, 982, 36, or 4, for example, VP3, or a sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto.

[0453] 457. Any one of the AAV particles from Embodiments 1 to 456, wherein the AAV capsid variant comprises an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with SEQ ID NO: 138.

[0454] 458. Any one of the AAV particles from Embodiments 1 to 457, wherein the AAV capsid variant comprises an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with SEQ ID NO: 138.

[0455] 459. The AAV capsid variant contains an amino acid sequence comprising at least 3, at least 4, at least 5, or at least 6 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941), (i) At least three consecutive amino acids contain SPH; (ii) At least four consecutive amino acids contain SPHS (SEQ ID NO: 4700); (iii) at least five consecutive amino acids containing SPHSK (SEQ ID NO: 4701); or (iv) At least six consecutive amino acids containing SPHSKA (SEQ ID NO: 941); The AAV capsid variant includes (a) a VP1 protein containing the amino acid sequence of SEQ ID NO: 981; (b) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 981; (c) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 981; or (d) an amino acid sequence having at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with any of the amino acid sequences in (a) to (c). One of the AAV particles from Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-458.

[0456] 460. The AAV capsid variant contains an amino acid sequence comprising at least 3, at least 4, at least 5, or at least 6 consecutive amino acids from the amino acid sequence of SPHSKA (SEQ ID NO: 941), (i) At least three consecutive amino acids contain SPH; (ii) At least four consecutive amino acids contain SPHS (SEQ ID NO: 4700); (iii) at least five consecutive amino acids containing SPHSK (SEQ ID NO: 4701); or (iv) At least six consecutive amino acids containing SPHSKA (SEQ ID NO: 941); The AAV capsid variant contains an amino acid sequence that is at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) of the amino acid sequence of SEQ ID NO: 981. One of the AAV particles from Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-459.

[0457] 461. The AAV capsid variant contains one or two but no more than three substitutions relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941), and the AAV capsid variant is (a) VP1 protein containing the amino acid sequence of SEQ ID NO: 981; (b) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 981; (c) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 981; or (d) An amino acid sequence having at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with any of the amino acid sequences in (a) to (c). including, One of the AAV particles from Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, or 446-460.

[0458] 462. Embodiments 1-23, 26-29, 32, 35-43, 46 in which the AAV capsid variant contains one or two but no more than three substitutions relative to the amino acid sequence of SPHSKA (SEQ ID NO: 941), and the AAV capsid variant contains an amino acid sequence that is at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 138 or SEQ ID NO: 981. Any one of the following AAV particles: ~51, 54~72, 69~89, 91, 94~99, 102~104, 107, 110~112, 115~129, 168~202, 240~247, 249, 250, 253~263, 266, 272~281, 286, 288, 291~299, 327~363, 369~371, 375~379, 385, 386, 390, 391, 395, 406, 408, 410~413, 415~432, 444, or 446~461.

[0459] 463. Any one of the AAV particles of Embodiments 459 to 462, wherein the amino acid sequence is located immediately after position 455, as numbered according to Sequence ID No. 138 or 981. 464. The AAV capsid variant contains an amino acid sequence comprising at least 3, at least 4, at least 5, or at least 6 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2), (i) At least three consecutive amino acids contain HDS; (ii) At least four consecutive amino acids contain HDSP (SEQ ID NO: 4702); (iii) at least five consecutive amino acids containing HDSPH (SEQ ID NO: 4703); or (iv) At least six consecutive amino acids comprising HDSPHK (SEQ ID NO: 2); The AAV capsid variant includes (a) a VP1 protein containing the amino acid sequence of SEQ ID NO: 982; (b) a VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 982; (c) a VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 982; or (d) an amino acid sequence having at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with any of the amino acid sequences in (a) to (c). Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264 Any one of the following AAV particles: ~280, 283~287, 289~299, 327~363, 369, 369, 371, 380~383, 385, 386, 390, 392, 395, 407, 409~412, 414, 433~435, 438~441, 443, 445, or 447~458.

[0460] 465. The AAV capsid variant contains an amino acid sequence comprising at least 3, at least 4, at least 5, or at least 6 consecutive amino acids from the amino acid sequence of HDSPHK (SEQ ID NO: 2), (i) At least three consecutive amino acids contain HDS; (ii) At least four consecutive amino acids contain HDSP (SEQ ID NO: 4702); (iii) at least five consecutive amino acids containing HDSPH (SEQ ID NO: 4703); or (iv) At least six consecutive amino acids comprising HDSPHK (SEQ ID NO: 2); The AAV capsid variant contains an amino acid sequence that is at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 982. Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-2 Any one of the following AAV particles: 80, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, or 464.

[0461] 466. The AAV capsid variant contains one or two but no more than three substitutions relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2), and the AAV capsid variant is (a) VP1 protein containing the amino acid sequence of SEQ ID NO: 982; (b) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 982; (c) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 982; or (d) An amino acid sequence having at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with any of the amino acid sequences in (a) to (c). including, Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280 , 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464, or 465 - one of these AAV particles.

[0462] 467. The AAV capsid variant contains one or two but no more than three substitutions relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2), and the AAV capsid variant contains an amino acid sequence that is at least 90% (e.g., at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) identical to the amino acid sequence of SEQ ID NO: 982, in embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52. One AAV particle from any of the following ranges: 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, or 464-466.

[0463] 468. An AAV particle of any one of embodiments 464 to 468, wherein the amino acid sequence is located immediately after position 453, as numbered according to SEQ ID NO: 138 or 982. 469. Any one of the AAV particles from Embodiments 1 to 468, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 981 or 982, or an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto.

[0464] 470. Any one of the AAV particles from Embodiments 1 to 469, wherein the AAV capsid variant comprises an amino acid sequence having at least one, at least two, or at least three modifications, e.g., substitutions (e.g., conservative substitutions), but 30 or fewer, 20 or fewer, or 10 or fewer modifications, e.g., substitutions (e.g., conservative substitutions) relative to the amino acid sequence of SEQ ID NO: 981 or 982.

[0465] 471. Any one of Embodiments 1 to 470, wherein the AAV capsid variant contains an amino acid sequence that includes at least one, at least two, or at least three different amino acids relative to the amino acid sequence of SEQ ID NO: 981 or 982, but with a total of 30 or fewer, 20 or fewer, or 10 or fewer amino acids.

[0466] 472. Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, in which the AAV capsid variant includes an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity with the amino acid sequence of SEQ ID NO: 981, or an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity therewith. , one of the following AAV particles: 102-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, or 469-471.

[0467] 473. The AAV capsid variant includes an amino acid sequence in which the amino acid sequence of SEQ ID NO: 981 has at least one, at least two, or at least three modifications, e.g., substitutions (e.g., conservative substitutions), but no more than 30, no more than 20, or no more than 10 modifications, e.g., substitutions (e.g., conservative substitutions), in embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 10 One AAV particle from any of the following ranges: 2-104, 107, 110-112, 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, or 469-472.

[0468] 474. The AAV capsid variant includes an amino acid sequence in which, with respect to the amino acid sequence of SEQ ID NO: 981, there are at least one, at least two, or at least three different amino acids, but no more than 30, no more than 20, or no more than 10 different amino acids, as defined in Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112. , one of the following AAV particles: 115-129, 168-202, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, or 469-473.

[0469] 475. The AAV capsid variant includes the amino acid sequence of SEQ ID NO: 982, or an amino acid sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity therewith, in embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90- Any one of the following AAV particles: 97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, or 464-471.

[0470] 476. Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 10, 10, 2 One AAV particle from any of the following ranges: 0-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, or 475.

[0471] 477. The AAV capsid variant includes an amino acid sequence in which, with respect to the amino acid sequence of SEQ ID NO: 982, there are at least one, at least two, or at least three different amino acids, but no more than 30, no more than 20, or no more than 10 different amino acids, as described in Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 1 One of the following AAV particles: 13-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, 475, or 476.

[0472] 478. Any one of the AAV particles from Embodiments 1 to 477, wherein the AAV capsid variant comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity therewith.

[0473] 479. Any one of Embodiments 1 to 478, wherein the nucleotide sequence encoding the capsid variant includes the nucleotide sequence of Sequence ID No. 983 or 984, or a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity therewith.

[0474] 480. Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94- Any one of the following AAV particles: 99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363, 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, 469-474, 478, or 479.

[0475] 481. Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, wherein the nucleotide sequence encoding the capsid variant includes the nucleotide sequence of SEQ ID NO: 984, or a nucleotide sequence having at least 80% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%) sequence identity thereto. , 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, or 475-479, one of these AAV particles.

[0476] 482. An AAV particle from any one of the preceding embodiments, in which the nucleotide sequence encoding a capsid variant is codon-optimized. 483. Embodiments 1-23, 26-29, 32, 35-43, 46-51, 54-72, 69-89, 91, 94-99, 102-104, 107, 110-112, 115-129, 168-202, 02, 240-247, 249, 250, 253-263, 266, 272-281, 286, 288, 291-299, 327-363 AAV particle comprising any one of the following amino acid sequences: 369-371, 375-379, 385, 386, 390, 391, 395, 406, 408, 410-413, 415-432, 444, 446-463, 469-474, 478-480, or 482, further comprising an amino acid sequence that is at least 95% identical to sequence number 981.

[0477] 484. Adeno-associated virus (AAV) particles comprising nucleic acids encoding an AAV capsid variant and a syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein), wherein the AAV capsid variant comprises the amino acid sequence of Sequence ID No. 981.

[0478] 485. An AAV particle of Embodiment 483 or 484, wherein the nucleotide sequence encoding the AAV capsid variant comprises the nucleotide sequence of Sequence ID No. 983, or a nucleotide sequence that is at least 90%, at least 95%, or at least 99% identical thereto.

[0479] 486. Embodiments 1-9, 11, 12-22, 24, 26, 28, 30, 33, 35-42, 44, 46-50, 52, 54-77, 83-88, 90-97, 100-103, 105, 110, 111, 113-129, 203-248, 251, 252, 254-257, 260, 261, 264-280, 283-287, 289-299, 327-363, 369, An AAV particle comprising any one of the following: 369, 371, 380-383, 385, 386, 390, 392, 395, 407, 409-412, 414, 433-435, 438-441, 443, 445, 447-458, 464-471, 475-479, 481, or 482, further comprising an amino acid sequence that is at least 95% identical to sequence number 982.

[0480] 487. An AAV particle comprising a nucleic acid encoding an AAV capsid variant and a syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein), wherein the AAV capsid variant comprises the amino acid sequence of Sequence ID No. 982.

[0481] 488. An AAV particle of Embodiment 486 or 487, wherein the nucleotide sequence encoding the AAV capsid variant comprises the nucleotide sequence of Sequence ID No. 984, or a nucleotide sequence that is at least 90%, at least 95%, or at least 99% identical thereto.

[0482] 489. Adeno-associated virus (AAV) particles comprising nucleic acids encoding an AAV capsid variant and a syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein), wherein the AAV capsid variant comprises an amino acid sequence encoded by the nucleotide sequence of SEQ ID NO: 983 or 984, or a nucleotide sequence that is at least 95% identical thereto.

[0483] 490. Adeno-associated virus (AAV) particles comprising nucleic acids encoding an AAV capsid variant and a syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein), wherein the AAV capsid variant comprises one amino acid sequence of SEQ ID NO: 4 or 36-59, and optionally, the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 4 or 36.

[0484] 491. Adeno-associated virus (AAV) particles comprising nucleic acids encoding an AAV capsid variant and a syntaxin-binding protein 1 (STXBP1) protein (e.g., human STXBP1 protein), wherein the AAV capsid variant comprises an amino acid sequence encoded by any one nucleotide sequence of sequence numbers 12-35, or a nucleotide sequence that is at least 95% identical thereto.

[0485] 492. AAV particle 490 or 491, wherein the nucleotide sequence encoding the AAV capsid variant contains one of the nucleotide sequences of sequence numbers 12-35, or a nucleotide sequence that is at least 95% identical thereto.

[0486] 493. Any one of Embodiments 1-299, 369-371, or 375-492 having increased directivity to CNS cells or tissues, such as brain cells, brain tissue, spinal cord cells, or spinal cord tissue, compared to AAV particles containing a capsid containing the amino acid sequence of SEQ ID NO: 138.

[0487] 494. Transducing a brain region, e.g., a midbrain region (e.g., hippocampus, or thalamus) or brainstem, and optionally measuring the level of transduction by an assay, e.g., an immunohistochemical assay or a qPCR assay, as described in Example 2, one of any AAV particles from Embodiments 1-129, 168-299, 369-371, or 375-493, where the level of transduction is at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, or at least 65 times greater compared to an AAV particle containing the capsid of SEQ ID NO: 138.

[0488] 495. Transducing a brain region, e.g., a midbrain region (e.g., hippocampus, or thalamus) or brainstem, and optionally measuring the level of transduction by an assay, e.g., an immunohistochemical assay or a qPCR assay, as described in Example 2, one of any AAV particles from Embodiments 1-129, 168-299, 369-371, or 375-494, where the level of transduction is at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, or at least 65 times greater compared to an AAV particle containing the capsid of SEQ ID NO: 138.

[0489] 496. Any one of the AAV particles from Embodiments 1-129, 168-299, 369-371, or 375-495, which, for example, are concentrated at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 times in the brain compared to the AAV particle containing the capsid of SEQ ID NO: 138, as measured by the assay described in Example 1.

[0490] 497. Any one of the AAV particles from Embodiments 1-129, 168-299, 369-371, or 375-496, which, for example, are concentrated at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, or at least 85 times in the brain compared to the AAV particles containing the capsid of SEQ ID NO: 138, as measured by the assay described in Example 1.

[0491] 498. For example, one AAV particle from any one of Embodiments 1-129, 168-299, 369-371, or 375-497, which is enriched in the brains of at least two to at least three species, e.g., non-human primates and rodents (e.g., mice), compared to an AAV particle containing the capsid of Sequence ID No. 138.

[0492] 499. For example, when measured by assay as described in Example 1 or 5, compared to AAV particles containing the capsid of SEQ ID NO: 138, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90 in the brains of at least 2 to at least 3 species, e.g., non-human primates and rodents (e.g., mice). AAV particles of any one of Embodiments 1-129, 168-299, 369-371, or 375-498, concentrated at least 95, at least 100, at least 105, at least 115, at least 120, at least 125, at least 130, at least 135, at least 140, at least 145, at least 150, at least 155, at least 160, at least 165, at least 170, at least 175, at least 180, at least 190, at least 200, at least 205, or at least 210 times.

[0493] 500. AAV particles of Embodiment 498 or 499, wherein at least two to at least three species are cynomolgus macaque (Macaca fascicularis), green macaque (Chlorocebus sabaeus), common marmoset (Callithrix jacchus), and / or mouse (e.g., BALB / c mouse, C57Bl / 6 mouse, and / or CD-1 uninbred mouse).

[0494] 501. Any one of the AAV particles from Embodiments 130-146, 369, 410-414, 447-454, 457, 458, 482, or 493, which are concentrated at least 2, at least 2.5, at least 3, at least 3.5, at least 4, at least 4.5, at least 5, at least 5.5, at least 6, at least 6.5, at least 7, at least 7.5, or at least 8 times in the brain compared to the AAV particle containing the capsid of SEQ ID NO: 981, as measured by the assay as described in Example 3.

[0495] 502. Any one of the AAV particles from Embodiments 147-167, 369, 410-414, 447-454, 457, 458, 482, or 493, which is concentrated at least 2, at least 2.5, at least 3, at least 3.5, at least 4, at least 4.5, at least 5, or at least 5.5 times in the brain compared to the AAV particle containing the capsid of SEQ ID NO: 982, as measured by the assay as described in Example 3.

[0496] 503. An AAV particle from any one of Embodiments 1-129, 168-299, 369-371, or 375-500, which delivers an increased level of STXBP1 to a brain region, and optionally, when the level of STXBP1 is measured by an assay, e.g., qRT-PCR or qPCR assay (e.g., as described in Example 2 or 8), it is increased by at least 10, at least 12, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, or at least 70 times compared to an AAV particle containing the capsid of SEQ ID NO: 138.

[0497] 504. An AAV particle from any one of Embodiments 1-129, 168-299, 369-371, 375-500, or 503, which delivers an augmented level of viral genome to a brain region, and optionally, when the level of viral genome is measured by an assay, e.g., qRT-PCR or qPCR assay (e.g., as described in Example 2 or 8), it is increased by at least 5, at least 10, at least 15, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, or at least 50-fold compared to an AAV particle containing the capsid of SEQ ID NO: 138.

[0498] 505. AAV particles of Embodiment 503 or 504, wherein the brain region is a midbrain region (e.g., hippocampus or thalamus), frontal cortex, temporal cortex, motor cortex, cerebral cortex, caudate nucleus, putamen, dentate nucleus, substantia nigra, or brainstem.

[0499] 506. For example, one AAV particle from any one of Embodiments 1-129, 168-299, 369-371, 375-500, or 503-505, which is concentrated at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, or at least 35 times in the spinal cord compared to an AAV particle containing the capsid of SEQ ID NO: 138, when measured by the assay as described in Example 1 or 8, and optionally the region of the spinal cord is the thoracic region, the cervical region, the C5 anterior horn region, the lumbar region, or the L5 anterior horn region.

[0500] 507. Any one of the AAV particles of Embodiments 1-129, 168-299, 369-371, 375-500, or 503-506, which exhibit preferential transduction in brain regions compared to transduction in the dorsal root ganglia (DRG).

[0501] 508. Any one of the AAV particles of Embodiments 1-129, 168-299, 369-371, 375-500, or 503-507, which exhibit preferential transduction in the brain region compared to the liver.

[0502] 509. Any one of the AAV particles of Embodiments 1-129, 168-299, 369-371, 375-500, or 503-508, which exhibit preferential transduction in brain regions compared to transduction in the heart.

[0503] 510. Any one of the AAV particles of Embodiments 1-129, 168-299, 369-371, 375-500, or 503-509, which exhibit preferential transduction in brain regions compared to transduction in the dorsal root ganglia (DRG) and cardiac regions.

[0504] 511. An AAV particle of any one of the preceding embodiments that can transduce non-neuronal cells, such as glial cells (e.g., oligodendrocytes or astrocytes).

[0505] 512. AAV particles of Embodiment 511, wherein the non-neuronal cells include glial cells, oligodendrocytes (e.g., Olig2-positive oligodendrocytes), or astrocytes (e.g., Olig2-positive astrocytes).

[0506] 513. AAV particles of any one of the preceding embodiments that can transduce Olig2-positive cells, such as Olig2-positive astrocytes or Olig2-positive oligodendrocytes.

[0507] 514. Any one of the AAV particles of Embodiments 369, 373, 447-454, 457, 458, or 482, having increased directivity toward cardiac cells or tissues, e.g., ventricles or atria, compared to the directivity of AAV particles containing the capsid of Sequence ID No. 138.

[0508] 515. Any one of the AAV particles of Embodiments 369, 373, 447-454, 457, 458, 482, or 514, which are concentrated at least 4, at least 5, at least 8, at least 10, at least 11, at least 12, at least 13, at least 14, at least 18, at least 19, at least 20, at least 21, at least 22, at least 24, at least 25, at least 27, at least 31, at least 33, or at least 34 times in the heart compared to the AAV particles containing the capsid of SEQ ID NO: 138, for example, as measured by the assay described in Example 4.

[0509] 516. Any one of the AAV particles of Embodiments 369, 374, 447-454, 457, 458, 482, having increased directivity to muscle cells or tissues (e.g., quadriceps muscle cells or quadriceps muscle tissue) compared to the directivity of AAV particles containing a capsid containing the amino acid sequence of Sequence ID No. 138.

[0510] 517. For example, one AAV particle from any one of Embodiments 369, 374, 447-454, 457, 458, 482, which is concentrated at least 4, at least 5, at least 8, at least 12, at least 17, at least 18, at least 20, at least 26, at least 27, at least 28, at least 30, or at least 36 times in muscle compared to an AAV particle containing the capsid of SEQ ID NO: 138, as measured by the assay described in Example 4.

[0511] 518. AAV particles of Embodiment 516 or 517, wherein the muscle cells or tissue are cardiac muscle (e.g., ventricle or atrium, or both), quadriceps muscle, or both. 519. An AAV particle of any one of the preceding embodiments, which is isolated and / or recombinant.

[0512] [Embodiments 520-585 are intentionally omitted.] 586. An AAV particle of any one of the preceding embodiments, comprising a promoter in which the viral genome is operably linked to the STXBP1 coding sequence.

[0513] 587. Promoters include human elongation factor 1α-subunit (EF1α), cytomegalovirus (CMV) early enhancer and / or promoter, chicken β-actin (CBA) promoter, CAG promoter, CAG derivative promoter, β-glucuronidase (GUSB) promoter, or ubiquitin C (UBC) promoter, neuron-specific enolase (NSE) promoter, platelet-derived growth factor (PDGF) promoter, platelet-derived growth factor B chain (PDGF-β) promoter, intercellular adhesion molecule 2 (ICAM-2) promoter, synapsin (Syn) promoter, methyl-CpG-binding protein 2 (MeCP2) promoter, and Ca2+ / calmodulin-dependent protein kinase II (CaMK II) AAV particles of Embodiment 586, which are metabotropic glutamate receptor 2 (mGluR2) promoters, neurofilament light chain (NFL) or heavy chain (NFH) promoters, β-globin minigene nβ2, preproenkephalin (PPE) promoters, enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2) promoters, glial fiber acidic protein (GFAP) promoters, myelin basic protein (MBP) promoters, cardiovascular promoters (e.g., αMHC, cTnT, and CMV-MLC2k), liver promoters (e.g., hAAT, TBG), skeletal muscle promoters (e.g., desmin, MCK, C512) or fragments thereof, e.g., shortened forms or functional variants.

[0514] [Embodiments 588 and 589 are intentionally omitted.] 590. An AAV particle from any one of embodiments 586 to 589, wherein the viral genome further comprises a polyA sequence.

[0515] 591. An AAV particle of any one of embodiments 586 to 590, wherein the viral genome further includes an inverted end sequence (ITR) sequence. 592. Any one of the AAV particles of Embodiments 586 to 591, wherein the viral genome includes an ITR sequence located 5' relative to the STXBP1 coding sequence (e.g., encoding the human STXBP1 protein).

[0516] 593. Any one of the AAV particles from Embodiments 586 to 592, wherein the viral genome includes an ITR sequence located 3' relative to the STXBP1 coding sequence (e.g., encoding the human STXBP1 protein).

[0517] 594. Any one of the AAV particles of Embodiments 586 to 593, wherein the viral genome includes an ITR sequence located at 5' relative to the STXBP1 coding sequence (e.g., encoding the human STXBP1 protein) and an ITR sequence located at 3' relative to the STXBP1 coding sequence (e.g., encoding the human STXBP1 protein).

[0518] 595. An AAV particle of any one of embodiments 586 to 594, wherein the viral genome further comprises enhancers, Kozak sequences, intron regions, and / or exon regions. [Embodiments 596-615 are intentionally omitted.] 616. Any one of the AAV particles of Embodiments 586 to 595, wherein the viral genome further comprises a nucleotide sequence encoding a Rep protein, e.g., a non-structural protein, and the Rep protein comprises Rep78 protein, Rep68 protein, Rep52 protein, and / or Rep40 protein (e.g., Rep78 and Rep52 proteins).

[0519] 617. AAV particles of Embodiment 616, wherein the AAV particles further comprise a nucleotide sequence encoding a Rep protein, e.g., a non-structural protein, and the Rep protein comprises a Rep78 protein, a Rep68 protein, a Rep52 protein, and / or a Rep40 protein (e.g., Rep78 and Rep52 proteins).

[0520] 618. AAV particles of Embodiment 616 or 617, wherein the Rep78 protein, Rep68 protein, Rep52 protein, and / or Rep40 protein are encoded by at least one Rep gene.

[0521] 619. Any one AAV particle of Embodiments 586 to 618, wherein the viral genome further comprises a nucleic acid sequence encoding an AAV capsid variant of any one AAV particle of Embodiments 1 to 519, 566, or 574.

[0522] 620. An AAV particle from any one of embodiments 575 to 619, wherein the AAV particle is an isolated and / or recombinant AAV particle. [Embodiment 621 is intentionally omitted.] 622. A cell containing an AAV particle of any one of the preceding embodiments, for example, a host cell.

[0523] 623. The cell of embodiment 622, wherein the cell is a mammalian cell or an insect cell. 624. Cells of Embodiment 622 or 623, wherein the cells are cells of a brain region or spinal cord region, optionally, cells of the brainstem, hippocampus, or thalamus.

[0524] 625. Any one of embodiments 622 to 624, wherein the cell is a neuron, sensory neuron, motor neuron, astrocyte, glial cell, oligodendrocyte, or muscle cell (e.g., heart, diaphragm, or quadriceps muscle cell).

[0525] [Embodiment 626 is intentionally omitted.] 627. A method for producing adeno-associated virus (AAV) particles, (i) to provide a host cell containing a viral genome including the STXBP1 coding sequence; and (ii) Incubate host cells under conditions suitable for encapsulating the viral genome in an AAV capsid variant as described in any one of Embodiments 1 to 620; This will allow us to create AAV particles. Methods that include...

[0526] 628. The method of Embodiment 627, further comprising introducing a first nucleic acid molecule containing a viral genome into a host cell prior to step (i). 629. The method of Embodiment 628, wherein the host cell contains a second nucleic acid encoding a capsid variant.

[0527] 630. The method of Embodiment 629, wherein the second nucleic acid molecule is introduced into the host cell before, simultaneously with, or after the first nucleic acid molecule. 631. A pharmaceutical composition comprising any one of the AAV particles of Embodiments 1 to 620 and a pharmaceutically acceptable excipient.

[0528] 632. A method for delivering a payload to a cell or tissue (e.g., a CNS cell or CNS tissue), comprising administering an effective amount of the pharmaceutical composition of Embodiment 631 or one of the AAV particles of Embodiments 1 to 620.

[0529] 633. The method of Embodiment 632, wherein the cells are cells from a brain region or spinal cord region, optionally selected from the prefrontal cortex, sensory cortex, motor cortex, caudate nucleus, cerebellar cortex, cerebral cortex, brainstem, hippocampus, or thalamus.

[0530] 634. The method of Embodiment 632 or 633, wherein the cells are neurons, sensory neurons, motor neurons, astrocytes, glial cells, or oligodendrocytes. [Embodiment 635 is intentionally omitted.] 636. Any one of embodiments 632 to 634, wherein cells or tissues are present within the subject.

[0531] 637. The method of Embodiment 636, wherein the subject has, has been diagnosed with, or is at risk of having, a genetic disorder, such as a monogenic disorder or a polygenic disorder.

[0532] 638. The method of Embodiment 636 or 637, wherein the subject has, has been diagnosed with, or is at risk of having, a neurological disorder, such as a neurodegenerative disorder. [Embodiment 639 is intentionally omitted.] 640. The method of Embodiment 636 or 637, wherein the subject has, has been diagnosed with, or is at risk of having, a muscle disorder or neuromuscular disorder.

[0533] 641. A method for treating a subject having or diagnosed with a genetic disorder, such as a monogenic disorder or a polygenic disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of Embodiment 631 or one of the AAV particles of Embodiments 1 to 620.

[0534] 642. A method for treating a subject having or diagnosed with a neurological disorder, such as a neurodegenerative disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of Embodiment 631 or one of the AAV particles of Embodiments 1 to 620.

[0535] 643. A method for treating a subject having or diagnosed with a muscle disorder or neuromuscular disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of Embodiment 631 or one of the AAV particles of Embodiments 1 to 620.

[0536] [Embodiment 644 is intentionally omitted.] 645. Any one of Embodiments 637 to 643, wherein the hereditary disorder, neurological disorder, neurodegenerative disorder, muscle disorder, or neuromuscular disorder is STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonic epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutation and further mutations by optional selection), Dravet syndrome (not caused by SCN1A mutation), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutation).

[0537] 646. Any one of embodiments 641 to 645, wherein treatment includes preventing the progression of a disability in the subject. 647. The methods of embodiments 636-646, wherein the subject is a human.

[0538] 648. Any one of Embodiments 636 to 647, wherein AAV particles are administered to a subject intravenously via intracisterna magna injection (ICM), intraparenchymally via intrathecal, intracerebroventricular, intraarterial, or intramuscular administration.

[0539] 649. Any one of embodiments 636 to 648, wherein AAV particles are administered to a subject, for example, via focused ultrasound (FUS-MB) conjugated to intravenous administration of microbubbles, or via MRI-guided FUS conjugated to intravenous administration.

[0540] 650. Any one of embodiments 636 to 649, wherein AAV particles are administered intravenously to a subject. 651. Any one of embodiments 636 to 650, wherein AAV particles are administered to a subject via intracisional injection (ICM).

[0541] 652. Any one of embodiments 636 to 651, wherein AAV particles are administered intra-arterially to a subject. [Embodiment 653 is intentionally omitted.] 654. Any one of embodiments 648 to 652, wherein administration of AAV particles results in an increase in the presence, level, and / or activity of the STXBP1 gene, mRNA, protein, or combination thereof.

[0542] 655. A pharmaceutical composition of Embodiment 631 or any one of Embodiments 1 to 620 for use in a method of delivering the STXBP1 coding sequence to cells or tissues. 656. A pharmaceutical composition of Embodiment 631 or any one of Embodiments 1 to 620 for use in a method of treating a genetic disorder, neurological disorder, neurodegenerative disorder, muscle disorder, or neuromuscular disorder.

[0543] 657. A pharmaceutical composition of Embodiment 631 or any one of Embodiments 1 to 620 for use in the manufacture of pharmaceuticals. 658. Use of the pharmaceutical composition of Embodiment 631 or any one of the AAV particles of Embodiments 1 to 620 in the manufacture of a pharmaceutical.

[0544] 659. Use of the pharmaceutical composition of Embodiment 631 or any one of the AAV particles of Embodiments 1 to 620 in the manufacture of a pharmaceutical for the treatment of a genetic disorder, neurological disorder, neurodegenerative disorder, muscle disorder, or neuromuscular disorder.

[0545] 660. An AAV particle comprising a viral genome comprising an AAV capsid variant of any one of Embodiments 1 to 519, for example, any one of Embodiments 507 to 510, and a nucleotide sequence encoding a syntaxin-binding protein 1 (STXBP1) protein (i.e., an STXBP1 coding sequence), wherein the nucleotide sequence 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 Sequence ID No. 6414.

[0546] 661. An AAV particle of Embodiment 660, wherein the nucleotide sequence encoding the STXBP1 protein (i.e., the STXBP1 coding sequence) contains a nucleotide sequence that is at least 90% identical to sequence number 6414.

[0547] 662. AAV particles of Embodiment 660 or Embodiment 661, wherein the nucleotide sequence encoding the STXBP1 protein (i.e., the STXBP1 coding sequence) contains a nucleotide sequence that is at least 95% identical to SEQ ID NO: 6414.

[0548] 663. Any one of embodiments 660 to 662, wherein the nucleotide sequence encoding the STXBP1 protein (i.e., the STXBP1 coding sequence) comprises the nucleotide sequence of sequence number 6414.

[0549] [Embodiment 664 is intentionally omitted.] 665. An AAV particle of any one of Embodiments 660 to 663, wherein the AAV capsid variant comprises (a) a VP1 protein containing the amino acid sequence of SEQ ID NO: 982; (b) a VP2 protein containing the amino acid sequence of positions 138 to 742 of SEQ ID NO: 982; or (c) a VP3 protein containing the amino acid sequence of positions 203 to 742 of SEQ ID NO: 982; or the AAV capsid variant is encoded by the nucleotide sequence of SEQ ID NO: 984 or a sequence that is at least 90% identical thereto (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).

[0550] 666. Any one of the AAV particles of Embodiments 660 to 663, wherein the AAV capsid variant contains three or fewer amino acid substitutions relative to the amino acid sequence of HDSPHK (SEQ ID NO: 2), and the AAV capsid variant contains an amino acid sequence that is at least 99% identical to SEQ ID NO: 982.

[0551] [Embodiments 667-668 are intentionally omitted.] 669. Any one of the AAV particles of Embodiments 660 to 666, wherein the encoded STXBP1 protein contains the amino acid sequence of Sequence ID No. 6413, or an amino acid 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 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.

[0552] 670. AAV particles of Embodiment 669, wherein the nucleotide sequence encoding the STXBP1 protein (i.e., the STXBP1 coding sequence) comprises the nucleotide sequence of Sequence ID No. 6414, 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.

[0553] 671. AAV particles of Embodiment 669 or 670, wherein the nucleotide sequence encoding the STXBP1 protein (i.e., the STXBP1 coding sequence) comprises the nucleotide sequence of Sequence ID No. 6414.

[0554] 672. An AAV particle of any one of embodiments 660 to 671, wherein the viral genome further includes a nucleotide sequence encoding a signal sequence. [Embodiment 673 is intentionally absent.]

[0555] 674. An AAV particle of Embodiment 672, wherein the nucleotide sequence encoding the signal sequence is located 5' relative to the STXBP1 coding sequence. 675. An AAV particle of Embodiment 674, wherein the STXBP1 coding sequence comprises the nucleotide sequence of Sequence ID No. 6414 or a nucleotide sequence that is at least 97% (e.g., at least 97%, at least 98%, or at least 99%) identical thereto.

[0556] 676. An AAV particle of Embodiment 675, wherein the STXBP1 coding sequence includes the nucleotide sequence of Sequence ID No. 6414. 677. An AAV particle of Embodiment 675, wherein the STXBP1 coding sequence consists of the nucleotide sequence of Sequence ID No. 6414.

[0557] 678. Any one of the AAV particles from embodiments 660 to 677, further comprising a promoter in which the viral genome is operably linked to the STXBP1 coding sequence. 679. An AAV particle of any one of embodiments 660 to 678, further comprising a viral genome and an enhancer.

[0558] [Embodiments 680-681 are intentionally omitted.] 682. AAV particles of Embodiment 678 or 679, wherein the promoter includes a tissue-specific promoter.

[0559] 683. AAV particles of Embodiment 678 or 679, wherein the promoter includes a ubiquitous promoter. 684. The promoter, EF-1a promoter, CBA promoter, chicken β-actin (CBA) promoter or derivative thereof, CAG promoter or derivative thereof, CMV early enhancer and / or promoter, β-glucuronidase (GUSB) promoter, ubiquitin C (UBC) promoter, neuron-specific enolase (NSE) promoter, platelet-derived growth factor (PDGF) promoter, platelet-derived growth factor B chain (PDGF-β) promoter, intercellular adhesion molecule 2 (ICAM-2) promoter, synapsin (Syn) promoter, synapsin 1 promoter (Syn1), methyl-CpG binding protein 2 (MeCP2) promoter, Ca2+ / calmodulin-dependent protein kinase II (CaMKII) promoter, A single AAV particle from any one of Embodiments 678 to 683, comprising a morphogenetic glutamate receptor 2 (mGluR2) promoter, a neurofilament light chain (NFL) or heavy chain (NFH) promoter, a β-globin minigene nβ2 promoter, a preproenkephalin (PPE) promoter, an enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2) promoter, a glial fiber acidic protein (GFAP) promoter, a myelin basic protein (MBP) promoter, a cardiovascular promoter (e.g., αMHC, cTnT, and CMV-MLC2k), a liver promoter (e.g., hAAT, TBG), a skeletal muscle promoter (e.g., desmin, MCK, C512) or a fragment thereof, e.g., a shortened form or a functional variant.

[0560] [Embodiments 685-712 are intentionally omitted.] 713. An AAV particle of any one of embodiments 660 to 684, wherein the viral genome further includes an inverted end sequence (ITR) sequence.

[0561] 714. An AAV particle of embodiment 713, wherein the ITR sequence is located 5' relative to the STXBP1 coding sequence. 715. An AAV particle of Embodiment 713 or Embodiment 714, wherein the ITR sequence is located 3' relative to the STXBP1 coding sequence.

[0562] 716. Any one of the AAV particles of Embodiments 713 to 715, wherein the viral genome includes an ITR located at 5' relative to the STXBP1 coding sequence and an ITR located at 3' relative to the STXBP1 coding sequence.

[0563] 722. An AAV particle of any one of embodiments 660 to 721, wherein the viral genome further includes a polyadenylated (poly-A) region. 724. An AAV particle of any one of embodiments 660 to 723, wherein the viral genome further includes introns.

[0564] 727. Any one of the AAV particles from Embodiments 660 to 726, wherein the viral genome further comprises exons, for example, at least one, at least two, or at least three exons.

[0565] 728. An AAV particle of any one of embodiments 660 to 727, wherein the viral genome further includes a Kozak sequence. [Embodiments 729-761 are intentionally omitted.] 762. An AAV particle of any one of embodiments 660 to 728, further comprising a nucleic acid encoding a Rep protein, wherein the Rep protein comprises a Rep78 protein, a Rep68 protein, a Rep52 protein, and / or a Rep40 protein.

[0566] 763. AAV particles of Embodiment 762, wherein the Rep78 protein, Rep68 protein, Rep52 protein, and / or Rep40 protein are encoded by at least one Rep gene.

[0567] 764. A vector encoding an AAV particle from any one of embodiments 660 to 763. 765. A cell containing an AAV particle of any one of embodiments 660 to 763 or a vector of embodiment 764.

[0568] 766. The cells of Embodiment 765, which are mammalian cells, e.g., HEK293 cells, insect cells, e.g., Sf9 cells, or bacterial cells. 767. A method for producing recombinant adeno-associated virus (AAV) particles, (i) To provide a host cell containing a viral genome that includes the nucleotide sequence of SEQ ID NO: 6414, or a sequence that is at least 90% (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) identical thereto; and (ii) Incubate host cells under conditions suitable for encapsulating the viral genome in a capsid variant containing the amino acid sequence of SEQ ID NO: 982; This will allow for the creation of recombinant AAV particles. Methods that include...

[0569] 768. The method of Embodiment 767, further comprising introducing a first nucleic acid molecule containing a viral genome into a host cell prior to step (i). 769. The method of Embodiment 767 or Embodiment 768, wherein the host cell contains a second nucleic acid encoding a capsid variant.

[0570] 770. The method of embodiment 769, further comprising introducing a second nucleic acid into cells. 771. The method of Embodiment 769 or Embodiment 770, wherein the second nucleic acid molecule is introduced into the host cell before, simultaneously with, or after the first nucleic acid molecule.

[0571] 772. A cell from any one of embodiments 767 to 772, wherein the host cell includes a mammalian cell, e.g., a HEK293 cell, an insect cell, e.g., an Sf9 cell, or a bacterial cell. 773. A pharmaceutical composition comprising any one of the AAV particles of Embodiments 660 to 763 and a pharmaceutically acceptable excipient.

[0572] 774. A method for delivering the STXBP1 protein to a target, comprising administering to the target an effective amount of the pharmaceutical composition of Embodiment 773 or one of the AAV particles from Embodiments 660 to 763, thereby delivering the nucleic acid encoding the STXBP1 protein (i.e., the STXBP1 coding sequence) to the target.

[0573] 775. The method of Embodiment 774, wherein the subject has, has been diagnosed with, or is at risk of having, an STXBP1-related disorder. 776. The method of Embodiment 774 or Embodiment 775, wherein the subject has, has been diagnosed with, or is at risk of having, a neurodegenerative disorder or neuromuscular disorder.

[0574] 777. A method for treating a subject having or diagnosed with an STXBP1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of Embodiment 773 or one of the AAV particles from Embodiments 660 to 763, thereby treating the STXBP1-related disorder in the subject.

[0575] 778. A method for treating a subject having or diagnosed with a neurodegenerative disorder or neuromuscular disorder, comprising administering to the subject an effective amount of the pharmaceutical composition of Embodiment 773 or one of the AAV particles from Embodiments 660 to 763, thereby treating the neurodegenerative disorder or neuromuscular disorder in the subject.

[0576] 779. Any one of embodiments 774 to 778, wherein the STXBP1-related disorder or neurodegenerative or neuromuscular disorder is STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutation and further mutations by optional selection), Dravet syndrome (not caused by SCN1A mutation), and Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutation).

[0577] 780. A method for treating a subject having or diagnosed with STXBP1 encephalopathy, comprising administering an effective amount of the pharmaceutical composition of Embodiment 773 or one of the AAV particles of Embodiments 660 to 763, thereby treating STXBP1 encephalopathy in the subject.

[0578] 781. The method of Embodiment 779 or Embodiment 780, wherein the subject has one or more mutations in STXBP1. 784. A method for treating a subject having or diagnosed with STXBP1 encephalopathy, comprising administering an effective amount of the pharmaceutical composition of Embodiment 773 or one of the AAV particles of Embodiments 660 to 763, thereby treating STXBP1 encephalopathy in the subject.

[0579] [Embodiments 785-788 are intentionally omitted.] 789. Any one of embodiments 774 to 784, wherein the subject has a reduced level of STXBP1 activity compared to a reference level.

[0580] 790. The method of Embodiment 789, wherein the reference level includes the level of STXBP1 activity in subjects without STXBP1-related disorders, neuromuscular disorders, and / or neurodegenerative disorders. 791. Any one of embodiments 777 to 790, wherein the treatment results in the recovery of at least one symptom of STXBP1-related disorder, neurodegenerative disorder, and / or neuromuscular disorder in the subject.

[0581] 792. The method of Embodiment 791, wherein at least one symptom of STXBP1-related disorder, neurodegenerative disorder, and / or neuromuscular disorder includes, for example, reduced STXBP1 activity in immune cells (e.g., macrophages), accumulation of glucocerebrosides and other glycolipids, accumulation of synuclein aggregates (e.g., Lewy bodies), developmental delay, progressive brain disease, progressive dementia, ataxia, myoclonus, oculomotor dysfunction, bulbar palsy, general weakness, tremors of the limbs, depression, hallucinations, cognitive decline, dystonia, or a combination thereof.

[0582] 793. Any one of the embodiments 774 to 792, wherein the subject is a human. 794. Any one of the embodiments 774 to 793, wherein the subject is young, for example, between the ages of 6 and 20.

[0583] 795. Any one of the embodiments 774 to 793, wherein the subject is an adult, for example, over 20 years of age. 796. Any one of the methods of Embodiments 774 to 795, wherein the subject has one or more mutations in the STXBP1 gene, STXBP1 mRNA, and / or STXBP1 protein.

[0584] 797. Any one of embodiments 774 to 796, wherein AAV particles are administered to a subject intravenously, intrathalamically (ITH) via intracerebral, intramuscularly, intrathecally, intraparenchymally (IVS-MB) via intravenous administration of microbubbles, or MRI-guided IVS-MB or intravenous administration via IVS-MB, or intracisional injection (ICM).

[0585] 798. A method according to any one of embodiments 774 to 796, wherein AAV particles are administered intravenously. 799. Any one of embodiments 774 to 796, wherein AAV particles are administered by intravenous injection, and optionally the intravenous injection is, for example, focused ultrasound (FUS) (FUS-MB) conjugated with intravenous administration of microbubbles, or MRI-guided FUS conjugated with intravenous administration.

[0586] 800. Any one of Embodiments 774 to 799, wherein AAV particles are administered to cells, tissues, or regions of the CNS, for example, regions of the brain or spinal cord, for example, parenchyma, cortex, substantia nigra, caudate nucleus, cerebellum, striatum, corpus callosum, cerebellum, brainstem caudate nucleus-putamen, thalamus, superior colliculus, spinal cord, or a combination thereof.

[0587] 801. Any one of Embodiments 774 to 800, wherein AAV particles are administered to at least two tissues or regions of the CNS (e.g., bilateral administration). 802. Any one of embodiments 774 to 799, wherein AAV particles are administered into cerebrospinal fluid, serum, or a combination thereof.

[0588] 803. Any one of embodiments 774 to 802, further comprising performing a blood test, performing an imaging test, collecting a CNS biopsy sample, collecting a tissue biopsy (e.g., a lung, liver, or spleen biopsy), collecting a blood or serum sample, or collecting an aqueous cerebrospinal fluid biopsy.

[0589] 804. A method of any one of Embodiments 774 to 803, further comprising evaluating, for example, the level of STXBP1 expression in a subject, for example, in the cells, tissues, or body fluids of the subject, e.g., measuring the level of STXBP1 gene, STXBP1 mRNA, and / or STXBP1 protein expression, wherein optionally the level of STXBP1 protein is measured by an assay described herein, e.g., ELISA, Western blotting, or immunohistochemical assay.

[0590] 805. The method of Embodiment 804, wherein the level of STXBP1 expression is measured before, during, or after treatment with AAV particles. 806. The method of Embodiment 804 or Embodiment 805, wherein the cells or tissues are cells or tissues of the central nervous system (e.g., parenchyma) or peripheral cells or tissues (e.g., liver, heart, and / or spleen).

[0591] 807. Any one of embodiments 774 to 806, wherein the administration results in an increase in the level of STXBP1 protein expression in the cells or tissue of the subject compared to a reference level, e.g., an untreated subject, e.g., a subject not administered AAV particles.

[0592] 808. Any one of Embodiments 774 to 807, further comprising evaluating, for example, the level of STXBP1 activity in a subject, for example, in the cells or tissue of the subject, and optionally, the level of STXBP1 activity being measured by an assay described herein.

[0593] 809. Administration is (i) the level of STXBP1 activity in the subject's cells, tissues (e.g., cells or tissues of the CNS, e.g., cortex, striatum, thalamus, cerebellum, and / or brainstem) and / or body fluids (e.g., CSF and / or serum) compared to a reference level, e.g., an untreated subject, e.g., a subject not administered AAV particles; (ii) the level of viral genome (VG) per cell in the target CNS tissue (e.g., cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord) compared to peripheral tissue (the level of VG per cell is at least 4 to at least 10 lower than the level in the CNS tissue, as measured, for example, by the assay described herein); and / or (iii) the level of STXBP1 mRNA expression in cells or tissues (e.g., cells or tissues of the CNS, e.g., cortex, thalamus, and / or brainstem) compared to a reference level, e.g., untreated (e.g., not administered AAV particles) or endogenous STXBP1 mRNA level, as measured by the assay described herein. Any one of embodiments 774 to 808, which results in an increase in at least one, at least two, or all of the above.

[0594] 810. Any one of embodiments 774 to 809 further comprising the administration of an additional therapeutic agent and / or therapy suitable for the treatment or prevention of diseases, neurodegenerative disorders, and / or neuromuscular disorders associated with STXBP1 expression.

[0595] 811. The method of Embodiment 810, wherein additional therapeutic agents and / or therapies include antiepileptic drugs (e.g., levetiracetam, phenobarbital, clobazam, topiramate), adrenocorticotropic hormone, or a combination thereof.

[0596] 812. A pharmaceutical composition of Embodiment 773 or any one of Embodiments 660 to 763 for use in the treatment of STXBP1-related disorders, neuromuscular disorders, and / or neurodegenerative disorders.

[0597] 813. Use in the manufacture of a pharmaceutical for the treatment of STXBP1-related disorders, neuromuscular disorders, and / or neurodegenerative disorders, an effective amount of the pharmaceutical composition of Embodiment 773 or any one of the AAV particles of Embodiments 660 to 763.

[0598] Details of various aspects or embodiments of this disclosure are described below. Other features, purposes, and advantages of this disclosure will become apparent from this specification and the claims. In this specification, the singular includes the plural unless the context clearly indicates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art of this disclosure. In case of any conflict, this specification shall prevail. [Brief explanation of the drawing]

[0599] [Figure 1] This shows the in vivo distribution (VG / cell) in the motor cortex, frontal cortex, putamen, substantia nigra, dentate nucleus, cervical spinal cord, anterior horn, DRG, liver, and heart of cynomolgus monkeys 28 days after IV injection of TTM-002.GBA_VG17-HA, AAV9.GBA_VG17-HA, or a solvent control. [Figure 2]This shows the mRNA expression of the GBA1 transgene in the motor cortex, frontal cortex, putamen, substantia nigra, dentate nucleus, cervical spinal cord, anterior horn, DRG, liver, and heart of cynomolgus monkeys 28 days after IV injection of TTM-002.GBA_VG17-HA, AAV9.GBA_VG17-HA, or a solvent control. [Figure 3A] This graph shows the percentage of HA-positive cells in the mouse cortex on the Y axis (percentage of cells transduced by the indicated capsid variant) at doses indicated on the X axis (from highest to lowest dose: 1e14 vg / kg, 3.2e13 vg / kg, 1e13 vg / kg, 3.2e12 vg / kg, or 1e12 vg / kg) at 28 days after intravenous administration of AAV particles containing TTM-002 or TTM-027 AAV capsid variants. [Figure 3B] This graph shows mRNA transgene expression for housekeeping genes in the mouse brain on the Y axis at doses shown on the X axis (from highest to lowest dose: 1e14 vg / kg, 3.2e13 vg / kg, 1e13 vg / kg, 3.2e12 vg / kg, or 1e12 vg / kg) 28 days after intravenous administration of AAV particles containing TTM-002 or TTM-027 AAV capsid variant. [Figure 4A] This graph shows the percentage of transdextrin cells (%HA+ cells) with HA+ nuclei as measured by nuclear H2B-HA staining and colocalization with hematoxylin in the indicated brain regions (temporal cortex, caudate nucleus, thalamus, or hippocampus) of African green monkeys. Measurements were taken 28 days after intravenous injection of AAV particles containing a TTM-002 capsid variant or AAV9 capsid control and an autocomplementary genome encoding HA-tagged histone 2B protein at a dose of 1e13 VG / kg. [Figure 4B]This graph shows the percentage of HA+ cells among cells positive for the indicated marker (NeuN+ neurons, SM311+ neurons, GFAP+ astrocytes, or Sox9+ astrocytes) in the indicated brain regions (temporal cortex, caudate nucleus, thalamus, or hippocampus) of African green monkeys. Measurements were taken 28 days after intravenous injection of AAV particles containing a self-complementary genome encoding the TTM-002 capsid variant and HA-tagged histone 2B protein at a dose of 1e13 VG / kg. The plotted data in Figures 4A–4B represent one section per monkey (n=2). Quantitative image analysis was performed on 1e3–1e5 cells according to region size. All P-values ​​were obtained from independent two-sided t-tests. [Figure 5A] Figure 5A shows a series of graphs illustrating the directivity of TTM-001 and TTM-002 to AAV9 control in the brain and liver of mice 28 days after intravenous administration at a dose of 1e13 VG / kg. Figure 5A shows viral genome (VG) / diploid genome (DG) in the brain for AAV9 control, TTM-001, or TTM-002; Figure 5B shows brain RNA (diploid vs. AAV9) for AAV9 control, TTM-001, or TTM-002; Figure 5C shows VG / DG in the liver for AAV9 control, TTM-001, or TTM-002; and Figure 5D shows liver RNA (diploid vs. AAV9) for AAV9 control, TTM-001, or TTM-002. Each data point represents an individual mouse, and all plotted values ​​represent mean ± SD (n=3). P-values ​​are obtained from independent two-tailed t-tests. [Figure 5B] Same as above. [Figure 5C] Same as above. [Figure 5D] Same as above. [Modes for carrying out the invention]

[0600] overview This specification describes compositions comprising AAV capsid variants containing a sequence encoding the STXBP1 protein, e.g., wild-type STXBP1 protein, e.g., wild-type human STXBP1 protein. In some embodiments, the disclosure provides methods for delivering AAV capsid variants containing a sequence encoding the STXBP1 protein to target cells or tissues. In some embodiments, the disclosure provides methods for delivering AAV capsid variants, thereby providing the STXBP1 protein, e.g., wild-type STXBP1 protein, e.g., wild-type human STXBP1 protein, to target cells or tissues. In some embodiments, the AAV capsid variants described herein have enhanced CNS directivity compared to other cells or tissues in the body, e.g., liver and / or DRGs.

[0601] AAVs have proven useful as biological tools 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 replication, and their relatively benign immunogenicity profile. Engineered adeno-associated virus (AAV) capsids with improved brain targeting would be an attractive solution to the limitations of CNS delivery. AAV-derived vectors are a promising tool for clinical gene transfer due to their non-pathogenic nature, low immunogenicity profile, low integration into the host genome, and long-term transgene expression in non-dividing cells. However, the transduction efficiency of naturally occurring AAVs in certain organs is too low for clinical use, and capsid neutralization with existing neutralizing antibodies may hinder treatment for most patients. For these reasons, considerable effort has been made to obtain capsid variants with enhanced properties. Of the many approaches tested to date, significant progress has been attributed to the directional evolution of AAV capsids using in vitro or in vivo selection of capsid variants generated by capsid sequence randomization using either error-prone PCR, shuffling of various parental serotypes, or insertion of a short peptide in a defined position with fully randomized properties.

[0602] The viral genome may be modified to contain a functional recombinant virus loaded or engineered to target a specific tissue and express or deliver STXBP1, or a minimal set of components for the assembly of viral particles. The viral genome may encode the STXBP1 protein, and viral particles containing the genome may be delivered to target cells, tissues, or organisms. In some embodiments, the genome encodes the human STXBP1 protein, e.g., the wild-type STXBP1 protein. In some embodiments, the target cells are CNS cells. In some embodiments, the target tissue is CNS tissue. In some embodiments, the target CNS tissue is brain tissue.

[0603] In some embodiments, the genome encodes the wild-type STXBP1 protein. In some embodiments, the genome includes, for example, a codon-optimized, CpG-depleted (e.g., CpG-depleted) nucleotide sequence encoding the wild-type STXBP1 protein, compared to the wild-type STXBP1 coding sequence. In some embodiments, the target cells are CNS cells. In some embodiments, the target tissue is CNS tissue. The target CNS tissue may be brain tissue. In some embodiments, the brain target includes the caudate nucleus, putamen, thalamus, superior colliculus, cortex, and corpus callosum.

[0604] Gene therapy presents an alternative approach to treating STXBP1-related disorders such as STXBP1-associated neurodegeneration or neuromuscular disorders (e.g., STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutations and further mutations by random selection), Dravet syndrome (not caused by SCN1A mutations), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutations). AAV presents several As a result of its advantageous features, it is commonly used in gene therapy approaches. Without being constrained by theory, in some embodiments, the AAV particles described herein could be used to preferentially administer and / or deliver the gene encoding the STXBP1 protein (e.g., human STXBP1 protein) to the CNS. In some embodiments, the AAV particles described herein could be used to preferentially administer and / or...

Claims

1. Adeno-associated virus (AAV) particles, a) The following formula: [N1]-[N2]-[N3] (in the formula, (i) By choice, [N1] includes X1, X2, and X3, where at least one of X1, X2, or X3 is G; (ii) [N2] contains the amino acid sequence of SPH; and (iii) [N3] comprises X4, X5, and X6, where at least one of X4, X5, or X6 is a basic amino acid. AAV capsid variants containing amino acid sequences having the following characteristics; and b) Viral genome containing the syntaxin-binding protein 1 (STXBP1) coding sequence Adeno-associated virus (AAV) particles, including those containing adeno-associated virus (AAV).

2. The AAV particle according to claim 1, wherein the amino acid sequence [N1]-[N2]-[N3] is present in the hypervariable loop IV of the AAV capsid variant.

3. The AAV particle according to claim 1 or claim 2, wherein the AAV capsid variant is an AAV9 capsid variant.

4. The AAV particle according to any one of claims 1 to 3, wherein [N1] comprises X1, X2, and X3, and at least one of X1, X2, or X3 is G.

5. AAV particle according to any one of claims 1 to 4, wherein [N2]-[N3] contains the amino acid sequence of SPHSKA (SEQ ID NO: 941).

6. Adeno-associated virus (AAV) particles containing a viral genome including the syntaxin-binding protein 1 (STXBP1) coding sequence and an AAV9 capsid variant including the amino acid sequence of SPHSKA (SEQ ID NO: 941).

7. The AAV particle according to claim 6, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is present in the hypervariable loop IV of the AAV9 capsid variant.

8. The AAV particle according to claim 6 or claim 7, wherein the amino acid sequence of SPHSKA (sequence number 941) is located immediately after the amino acid position corresponding to position 455 of sequence number 4 or sequence number 36.

9. The AAV particle according to any one of claims 6 to 8, wherein the AAV9 capsid variant further comprises one, two, or all of the following: N at the amino acid position corresponding to position 452 of SEQ ID NO: 4, E at the amino acid position corresponding to position 451, and / or V at the amino acid position corresponding to position 453.

10. The AAV particle according to any one of claims 6 to 9, wherein the AAV9 capsid variant comprises the amino acid sequence KTENVSGSPHSKAQNQQT (SEQ ID NO: 3272).

11. The AAV9 capsid variant is (i) VP1 protein containing an amino acid sequence having at least 90% identity with SEQ ID NO: 4; (ii) A VP2 protein comprising an amino acid sequence having at least 90% identity with positions 138-742 of Sequence ID No. 4; and / or (iii) VP3 protein containing an amino acid sequence that has at least 90% identity with positions 203-742 of Sequence ID No. 4 AAV particles according to any one of claims 6 to 10, including the AAV particles described in any one of claims 6 to 10.

12. The AAV9 capsid variant is (i) VP1 protein containing an amino acid sequence having at least 95% identity with SEQ ID NO: 4; (ii) A VP2 protein comprising an amino acid sequence having at least 95% identity with positions 138-742 of Sequence ID No. 4; and / or (iii) VP3 protein containing an amino acid sequence that has at least 95% identity with positions 203-742 of Sequence ID No. 4 AAV particles according to any one of claims 6 to 11, including the AAV particles described in any one of claims 6 to 11.

13. The AAV9 capsid variant is (i) VP1 protein containing an amino acid sequence that is at least 99% identical to SEQ ID NO: 4; (ii) A VP2 protein comprising an amino acid sequence having at least 99% identity with positions 138-742 of Sequence ID No. 4; and / or (iii) VP3 protein containing an amino acid sequence that has at least 99% identity with positions 203-742 of Sequence ID No. 4 AAV particles according to any one of claims 6 to 12, including the AAV particles described in any one of claims 6 to 12.

14. The AAV9 capsid variant is (i) VP1 protein containing the amino acid sequence of Sequence ID No. 4; (ii) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 4; and / or (iii) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 4 AAV particles according to any one of claims 6 to 13, including the AAV particles described in any one of claims 6 to 13.

15. The AAV9 capsid variant is (i) The amino acid sequence of SPHSKA (SEQ ID NO: 941) (the amino acid sequence is located immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 4); (ii) E at the amino acid position corresponding to position 451 of sequence number 4 and V at the amino acid position corresponding to position 453. Includes; and (iii) Other modifications to the wild AAV9 AAV particles according to any one of claims 6 to 13, which do not contain the above.

16. The AAV particle according to any one of claims 6 to 8, wherein the AAV9 capsid variant further comprises one, two, or all of the following: E ​​at the amino acid position corresponding to position 451 of SEQ ID NO: 36, R at the amino acid position corresponding to position 452, and / or V at the amino acid position corresponding to position 453.

17. The AAV particle according to any one of claims 6 to 8 and 16, wherein the AAV9 capsid variant comprises the amino acid sequence KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).

18. The AAV9 capsid variant is (i) VP1 protein containing an amino acid sequence having at least 90% identity with SEQ ID NO: 36; (ii) A VP2 protein having an amino acid sequence that is at least 90% identical to positions 138-742 of Sequence ID No. 36; and / or (iii) VP3 protein containing an amino acid sequence that has at least 90% identity with positions 203-742 of Sequence ID No. 36 AAV particles according to any one of claims 6 to 8, 16, and 17, comprising:

19. The AAV9 capsid variant is (i) VP1 protein containing an amino acid sequence having at least 95% identity with SEQ ID NO: 36; (ii) A VP2 protein comprising an amino acid sequence having at least 95% identity with positions 138-742 of Sequence ID No. 36; and / or (iii) VP3 protein containing an amino acid sequence that has at least 95% identity with positions 203-742 of Sequence ID No. 36 AAV particles according to any one of claims 6 to 8 and 16 to 18, including the AAV particles described in any one of claims 6 to 8 and 16 to 18.

20. The AAV9 capsid variant is (i) VP1 protein containing an amino acid sequence that is at least 99% identical to SEQ ID NO: 36; (ii) A VP2 protein having an amino acid sequence that is at least 99% identical to positions 138-742 of Sequence ID No. 36; and / or (iii) VP3 protein containing an amino acid sequence that has at least 99% identity with positions 203-742 of Sequence ID No. 36 AAV particles according to any one of claims 6 to 8 and 16 to 19, including the AAV particles described in any one of claims 6 to 8 and 16 to 19.

21. The AAV9 capsid variant is (i) VP1 protein containing the amino acid sequence of SEQ ID NO: 36; (ii) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 36; and / or (iii) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 36 AAV particles according to any one of claims 6 to 8 and 16 to 20, including the AAV particles described in any one of claims 6 to 8 and 16 to 20.

22. The AAV9 capsid variant is (i) Amino acid sequence SPHSKA (SEQ ID NO: 941) (The above amino acid sequence is located immediately after the amino acid position corresponding to position 455 of SEQ ID NO: 36); (ii) E at the amino acid position corresponding to position 451 of sequence number 36, R at the amino acid position corresponding to position 452, and V at the amino acid position corresponding to position 453. Includes; and (iii) Other modifications to the wild AAV9 AAV particles according to any one of claims 6 to 8 and 16 to 20, which do not contain any of the above.

23. [N1]-[N2]-[N3] is located immediately after the position corresponding to amino acid position 452 of sequence number 982; and The AAV particle according to any one of claims 1 to 4, wherein the AAV capsid variant includes an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 982, for example, positions 203 to 742 of SEQ ID NO: 982, for example, 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.

24. The AAV particle according to claim 23, wherein [N1] contains GHD.

25. The AAV particle according to claim 23 or claim 24, wherein [N1] contains amino acid G at the position corresponding to position 453 of SEQ ID NO: 138 or SEQ ID NO: 982, amino acid H at position 454, and amino acid D at position 455.

26. AAV particle according to any one of claims 23 to 25, wherein [N3] contains KSG.

27. The AAV capsid variant is (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with SEQ ID NO: 982; (ii) A VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 982 AAV particles according to any one of claims 23 to 26, including the AAV particles described in any one of claims 23 to 26.

28. The AAV capsid variant is (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with SEQ ID NO: 982; (ii) A VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity with positions 203-742 of SEQ ID NO: 982 AAV particles according to any one of claims 23 to 27, including the AAV particles described in any one of claims 23 to 27.

29. The AAV capsid variant is (i) VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with SEQ ID NO: 982; (ii) A VP2 protein containing the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 138-742 of SEQ ID NO: 982; or (iii) VP3 protein containing the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity with positions 203-742 of SEQ ID NO: 982 AAV particles according to any one of claims 23 to 28, including the AAV particles described in any one of claims 23 to 28.

30. The AAV capsid variant is (i) VP1 protein containing the amino acid sequence of SEQ ID NO: 982; (ii) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 982; or (iii) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 982 AAV particles according to any one of claims 23 to 29, including the AAV particles described in any one of claims 23 to 29.

31. The AAV particle according to any one of claims 1 to 30, wherein the viral genome encodes the wild-type STXBP1 protein.

32. The AAV particle according to any one of claims 1 to 31, wherein the viral genome encodes the human STXBP1 protein.

33. The AAV particle according to claim 31 or claim 32, wherein the STXBP1 protein comprises the amino acid sequence of SEQ ID NO: 6413.

34. The AAV particle according to any one of claims 1 to 33, wherein the STXBP1 coding sequence includes a nucleotide sequence that is at least 90% identical to sequence number 6414 (for example, 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).

35. The AAV particle according to claim 34, wherein the STXBP1 coding sequence includes a nucleotide sequence that is at least 95% identical to sequence number 6414.

36. The AAV particle according to claim 35, wherein the STXBP1 coding sequence includes a nucleotide sequence that is at least 99% identical to sequence number 6414.

37. The AAV particle according to claim 36, wherein the STXBP1 coding sequence includes the nucleotide sequence of sequence number 6414.

38. The AAV particle according to claim 36, wherein the STXBP1 coding sequence consists of the nucleotide sequence of sequence number 6414.

39. The AAV particle according to any one of claims 1 to 38, wherein the viral genome comprises a promoter operably linked to the STXBP1 coding sequence.

40. The promoter is human elongation factor 1α-subunit (EF1α), cytomegalovirus (CMV) early enhancer and / or promoter, chicken β-actin (CBA), CAG, CAG derivatives, β-glucuronidase (GUSB), or ubiquitin C (UBC), neuron-specific enolase (NSE), platelet-derived growth factor (PDGF), platelet-derived growth factor B chain (PDGF-β), intercellular adhesion molecule 2 (ICAM-2), synapsin (Syn), synapsin 1 (Syn1), methyl-CpG-binding protein 2 (MeCP2), Ca2+ / calmodulin-dependent protein kinase II (CaMKII), metabotropic glutamate receptor 2 (mGluR2), neurofilament light chain (NFL). AAV particles according to claim 39, which are heavy chain (NFH), β-globin minigene nβ2, preproenkephalin (PPE), enkephalin (Enk), and excitatory amino acid transporter 2 (EAAT2), glial fiber acidic protein (GFAP), myelin basic protein (MBP), cardiovascular promoters (e.g., αMHC, cTnT, and CMV-MLC2k), liver promoters (e.g., hAAT, TBG), skeletal muscle promoters (e.g., desmin, MCK, C512) or fragments thereof, e.g., shortened forms or functional variants.

41. The AAV particle according to any one of claims 1 to 40, wherein the viral genome further comprises an inverted terminal sequence (ITR) sequence.

42. The AAV particle according to claim 41, wherein the viral genome includes an ITR sequence located at 5' with respect to the STXBP1 coding sequence.

43. The AAV particle according to claim 41 or claim 42, wherein the viral genome includes an ITR sequence located at 3' relative to the STXBP1 coding sequence.

44. The AAV particle according to any one of claims 41 to 43, wherein the viral genome includes an ITR sequence located at 5' with respect to the STXBP1 coding sequence and an ITR sequence located at 3' with respect to the STXBP1 coding sequence.

45. Viral genome including the syntaxin-binding protein 1 (STXBP1) coding sequence, and (i) VP1 protein containing the amino acid sequence of Sequence ID No. 4; (ii) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 4; and / or (iii) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 4 Adeno-associated virus (AAV) particles containing AAV capsid variants.

46. Viral genome including the syntaxin-binding protein 1 (STXBP1) coding sequence, and (i) VP1 protein containing the amino acid sequence of SEQ ID NO: 36; (ii) VP2 protein containing the amino acid sequence from positions 138 to 742 of SEQ ID NO: 36; and / or (iii) VP3 protein containing the amino acid sequence from positions 203 to 742 of SEQ ID NO: 36 Adeno-associated virus (AAV) particles containing AAV capsid variants.

47. A cell comprising AAV particles according to any one of claims 1 to 46, wherein the cell is optionally a mammalian cell (e.g., HEK293 cell), an insect cell (e.g., Sf9 cell), or a bacterial cell.

48. A method for producing AAV particles according to any one of claims 1 to 46, (i) To provide a cell comprising the viral genome comprising the STXBP1 coding sequence and the nucleic acid encoding the AAV capsid variant; and (ii) Incubate the cells under conditions suitable for encapsulating the viral genome in the AAV capsid variant; This is how the AAV particles are produced. Methods that include...

49. (a) The viral genome includes the nucleotide sequence of Sequence ID No. 6414 or a nucleotide sequence that is at least 90% identical thereto (for example, 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); and (b) The AAV capsid variant is (i) The amino acid sequence of SEQ ID NO: 4 or the VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 4 (for example, 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% identity); (ii) A VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 4 (for example, 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% identity); or (iii) A VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 4 (for example, 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% identity). The method according to claim 48, including the method described in claim 48.

50. The method according to claim 49, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 4, the amino acid sequence from positions 138 to 742 of SEQ ID NO: 4, and / or the amino acids from positions 203 to 742 of SEQ ID NO:

4.

51. (a) The viral genome includes the nucleotide sequence of Sequence ID No. 6414 or a nucleotide sequence that is at least 90% identical thereto (for example, 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); and (b) The AAV capsid variant is (i) The amino acid sequence of SEQ ID NO: 36 or a VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 36 (for example, 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% identity); (ii) A VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 36 (for example, 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% identity); or (iii) A VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 36 (for example, 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% identity). The method according to claim 48, including the method described in claim 48.

52. The method according to claim 51, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 36, the amino acid sequence from positions 138 to 742 of SEQ ID NO: 36, and / or the amino acids from positions 203 to 742 of SEQ ID NO:

36.

53. (a) The viral genome includes the nucleotide sequence of Sequence ID No. 6414 or a nucleotide sequence that is at least 90% identical thereto (for example, 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); and (b) The AAV capsid variant is (i) The amino acid sequence of SEQ ID NO: 982 or the VP1 protein comprising an amino acid sequence having at least 90% identity with SEQ ID NO: 982 (for example, 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% identity); (ii) A VP2 protein comprising the amino acid sequence at positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 138-742 of SEQ ID NO: 982 (for example, 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% identity); or (iii) A VP3 protein comprising the amino acid sequence at positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity with positions 203-742 of SEQ ID NO: 982 (for example, 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% identity). The method according to claim 48, including the method described in claim 48.

54. The method according to claim 53, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 982, the amino acid sequence from positions 138 to 742 of SEQ ID NO: 982, and / or the amino acid sequence from positions 203 to 742 of SEQ ID NO:

982.

55. The method according to any one of claims 48 to 54, further comprising introducing a first nucleic acid molecule containing the viral genome into the cell prior to step (i).

56. The method according to any one of claims 48 to 55, wherein the cells contain a second nucleic acid molecule encoding the AAV capsid variant, and optionally the method further comprises introducing the second nucleic acid molecule into the cells prior to step (i).

57. The method according to any one of claims 48 to 56, wherein the cells include mammalian cells (e.g., HEK293 cells), insect cells (e.g., Sf9 cells), or bacterial cells.

58. A pharmaceutical composition comprising AAV particles according to any one of claims 1 to 46 and a pharmaceutically acceptable excipient.

59. A pharmaceutical composition comprising AAV particles according to any one of claims 5 to 22 and a pharmaceutically acceptable excipient.

60. A pharmaceutical composition comprising AAV particles according to any one of claims 9 to 15 and 45, and a pharmaceutically acceptable excipient.

61. A pharmaceutical composition comprising AAV particles according to any one of claims 16 to 22 and 46, and a pharmaceutically acceptable excipient.

62. A method for delivering AAV particles encoding the STXBP1 protein to a target, comprising administering to the target an effective amount of the pharmaceutical composition according to any one of claims 58 to 61 or the AAV particles according to any one of claims 1 to 46.

63. The method according to claim 62, wherein the subject has, has been diagnosed with, or is at risk of having, an STXBP1-related disorder, and optionally the STXBP1-related disorder is STXBP1-related neurodegeneration or neuromuscular disorder.

64. The method according to claim 62 or 63, wherein the subject has, has been diagnosed with, or is at risk of having, STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonic epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutation and further mutations by optional selection), Dravet syndrome (not caused by SCN1A mutation), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutation).

65. A method for treating a subject having or diagnosed with an STXBP1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition according to any one of claims 58 to 61 or the AAV particles according to any one of claims 1 to 46.

66. A method for treating a subject having or diagnosed with an STXBP1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 59 or the AAV particles according to any one of claims 5 to 22.

67. A method for treating a subject having or diagnosed with an STXBP1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 60 or AAV particles according to any one of claims 9 to 15 and 45.

68. A method for treating a subject having or diagnosed with an STXBP1-related disorder, comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 61 or AAV particles according to any one of claims 16 to 22 and 46.

69. The method according to any one of claims 65 to 68, wherein the STXBP1-related disorder is STXBP1-related neurodegeneration or neuromuscular disorder.

70. The method according to claim 69, wherein the STXBP1-related neurodegeneration or neuromuscular disorder is STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutation and further mutations by optional selection), Dravet syndrome (not caused by SCN1A mutation), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutation).

71. Having STXBP1 encephalopathy or STXBP A method for treating a subject diagnosed with encephalopathy, comprising administering to the subject an effective amount of the pharmaceutical composition according to any one of claims 58 to 61 or the AAV particles according to any one of claims 1 to 46.

72. Having STXBP1 encephalopathy or STXBP A method for treating a subject diagnosed with 1. encephalopathy, comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 59 or AAV particles according to any one of claims 5 to 22.

73. Having STXBP1 encephalopathy or STXBP A method for treating a subject diagnosed with 1. encephalopathy, comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 60 or AAV particles according to any one of claims 9 to 15 and 45.

74. Having STXBP1 encephalopathy or STXBP A method for treating a subject diagnosed with 1. encephalopathy, comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 61 or AAV particles according to any one of claims 16 to 22 and 46.

75. The method according to any one of claims 62 to 74, wherein the subject has one or more mutations in the STXBP1 gene.

76. The method according to any one of claims 62 to 75, wherein the subject has a reduced level of STXBP1 activity compared to a reference level in a subject that does not have STXBP1-related impairment.

77. The method according to any one of claims 65 to 76, wherein the treatment results in the prevention of the progression of the disorder in the subject.

78. The method according to any one of claims 65 to 77, wherein the treatment results in the recovery of at least one symptom of the disorder and / or a change in one or more biomarkers of the disorder.

79. The method according to claim 78, wherein the one or more biomarkers include STXBP1 activity or neurofilament light chains.

80. The method according to claim 78, wherein the at least one symptom includes epilepsy, autistic features, ataxia, generalized tremor, dystonia, or a combination thereof.

81. The method according to any one of claims 62 to 80, wherein the subject is a human.

82. The method according to any one of claims 62 to 81, wherein the AAV particles are delivered to cells, tissues, or regions of the CNS, for example, regions of the brain or spinal cord, for example, parenchyma, cortex, substantia nigra, caudate nucleus-cerebellum, striatum, corpus callosum, cerebellum, brainstem, caudate nucleus-putamen, thalamus, superior colliculus, spinal cord, or a combination thereof, and / or neurons, for example, GABAergic neurons, glutamatergic neurons, or a combination thereof.

83. The method according to any one of claims 62 to 82, further comprising evaluating, for example, measuring, the level of STXBP1 expression in the subject, for example, cells, tissues, or body fluids of the subject, such as STXBP1 gene expression, STXBP1 mRNA expression, and / or STXBP1 protein expression.

84. The method according to claim 83, wherein the level of STXBP1 protein expression is measured by ELISA, Western blot, or immunohistochemical assay.

85. The method according to claim 83 or claim 84, wherein the level of STXBP1 expression is evaluated before and after administration of the AAV particles, and optionally, the level of STXBP1 expression before treatment is compared with the level of STXBP1 expression after administration.

86. The method according to any one of claims 83 to 85, comprising evaluating the level of STXBP1 expression in cells or tissues of the central nervous system (e.g., parenchyma).

87. The method according to any one of claims 83 to 86, wherein the level of expression of the target STXBP1 protein after administration is increased compared to the level of expression of the target STXBP1 protein before administration.

88. The method according to any one of claims 62 to 87, further comprising, for example, evaluating, for example, the level of STXBP1 activity in the cells or tissues of the subject, for example, measuring it.

89. The aforementioned administration, (i) STXBP1 activity in the cells, tissues (e.g., cells or tissues of the CNS, e.g., cortex, striatum, thalamus, cerebellum, and / or brainstem), and / or body fluids (e.g., CSF and / or serum) of the subject, compared to the STXBP1 activity in the subject before administration; (ii) The level of viral genome (VG) per cell in the CNS tissue of the subject (e.g., cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord) compared to the level of VG per cell in the peripheral tissue of the subject; and / or (iii) STXBP1 mRNA expression in the cells or tissues of the subject (e.g., cells or tissues of the CNS, e.g., cortex, thalamus, and / or brainstem) compared to the STXBP1 mRNA expression in the subject before administration. The method according to any one of claims 62 to 88, which results in an increase.

90. The aforementioned subjects further include administering additional agents suitable for the treatment or prevention of STXBP1-related disorders, The method according to any one of claims 65 to 89, wherein the additional agent optionally includes one or more antiepileptic drugs (e.g., levetiracetam, phenobarbital, clobazam, topiramate), adrenocorticotropic hormone, or a combination thereof.

91. The method according to any one of claims 65 to 90, further comprising administering an immunosuppressant to the subject.

92. The method according to claim 91, wherein the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxychloroquine.

93. A pharmaceutical composition according to any one of claims 58 to 61 or AAV particles according to any one of claims 1 to 46 for use in the treatment of STXBP1-related disorders; optionally, the STXBP1-related disorder is STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutation and optionally further mutations), Dravet syndrome (not caused by SCN1A mutation), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutation).

94. Use of a pharmaceutical composition according to any one of claims 58 to 61 or AAV particles according to any one of claims 1 to 46 in the manufacture of a pharmaceutical for the treatment of STXBP1-related disorders; optionally, the STXBP1-related disorder is STXBP1 encephalopathy, epileptic encephalopathy, Ohtahara syndrome, developmental encephalopathy, West syndrome, early myoclonus epileptic encephalopathy, Lennox-Gastaut syndrome, autism (e.g., autism with STXBP1 mutation and optionally further mutations), Dravet syndrome (not caused by SCN1A mutation), or Rett syndrome phenotype (not caused by MECP2 or CDKL5 mutation).

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