Compositions comprising peptides and methods of using the same with aav

Abstract

Disclosed herein are compositions comprising J peptides and methods of using the same to load a cargo into an AAV particle or an AAV-like particle.

Description

COMPOSITIONS COMPRISING PEPTIDES AND METHODS OF USING THE SAME WITH AAVI. CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 559,349 filed 29 February 2024, which is incorporated herein in its entirety.II. REFERENCE TO THE SEQUENCE LISTING

[0002] The Sequence Listing submitted 28 February 2025 as a text file named “24-3011- WO_SL”, created on 28 February 2025 and having a size of 233,472 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.52(e)(5).III. BACKGROUND

[0003] Adeno-associated virus (AAV) vectors are the leading platform for gene delivery for the treatment of a variety of human diseases. Recent advances in developing clinically desirable AAV capsids, optimizing genome designs and harnessing revolutionary biotechnologies have contributed substantially to the growth of the gene therapy field. Preclinical and clinical successes in AAV-mediated gene replacement, gene silencing and gene editing have helped AAV gain popularity as the ideal therapeutic vector, with several AAV-based therapeutics gaining regulatory approval in Europe and the United States. Continued study of AAV biology and increased understanding of the associated therapeutic challenges and limitations are necessary to build the foundation for future clinical success. To improve AAV gene therapy, researchers are focusing on optimizing the capsid design through engineering techniques like directed evolution and rational design, enhancing the transgene expression cassette by codon optimization and promoter selection, and developing strategies to target specific cell types by modifying the capsid to improve tissue tropism; all while aiming to minimize immune responses and maximize therapeutic efficacy in the target tissue

[0004] Thus, there remains an urgent need to develop AAV-based platforms to treat various genetic diseases and disorders. Consequently, the present disclosure provides compositions for and methods of using an inventive AAV-based platform alone or in combination with other treatments.IV. BRIEF DESCRIPTION OF THE FIGURES

[0005] FIG. 1 shows in silico solutions from 4,000 peptide predictions. The y-axis is predicted aligned error (PAE), where a smaller PAE implies better binding. Scores that are less than 10 are taken as validated in silico. Manual screening determined 20 / 49 solutions were sterically impossible in the context of a capsid but not a monomer. Predictions were made by RF Diffusion for backbone, filled in with MPNN, and PAE estimated by Alphafold2 from dl binder design. Solutions were named by [job].[RF Diffusion solution], [MPNN solution],

[0006] FIG. 2 shows in silico solutions from 15 peptide predictions that were screened by purification. The y axis is predicted aligned error (PAE), where a smaller PAE implies better binding. Scores that are less than 10 are taken as validated in silico.

[0007] FIG. 3 shows an example of a candidate predicted 5 times by Alphafold2 maintains a robust and consistent fold in silico.

[0008] FIG. 4A - FIG. 4C show J peptides (example green) were targeted against the beta strand G (purple) of the capsid. FIG. 4A shows space filled view of capsid interior with candidate J peptide. FIG. 4B shows a ribbon diagram of candidate J peptide with only |3G of the capsid shown. Secondary structure resulted in numerous hydrogen bond contacts often by inclusion of a novel P strand J, extending the beta sheet. FIG. 4C shows a one-dimensional depiction of J peptide candidate, where a predicted disulfide is depicted with a yellow line, and residues pointed into the buried AAV capsid lumin are demarked with and asterisk. Hydrogen bonds are predicted between strands PG and pj. Beta strands are depicted as arrows (red in J peptide, purple for capsid), while the alpha helix is depicted as a red cylinder. Structure taken from the AlphaFold2 prediction used to validate binders.

[0009] FIG. 5 shows an example of a candidate J peptide (blue) fused to cargo EGFP (green). The structure was generated with Alpahfold2.

[0010] FIG. 6 shows that AAV9 particles purified by affinity antibody also purified EGFP fused to candidate J peptides. Particles were produced by triple transfection method (XX680 helper, pLH9 (Rep2 / Cap9), and EFla-EGFP-J peptide fusion). Virus was harvested after 3 days from combined media / cells and freeze thawed to lyse, the cell pellet was clarified by centrifugation. Purification was done by AAVX resin in 10 pL reactions, the bead volume resuspended in RIP A buffer and run on SDS-PAGE gel. The candidates further evaluated are outlined in Red boxes.

[0011] FIG. 7A provides an alignment of J peptides that show that J peptides have little sequence conservation. FIG. 7B shows that solutions sharing job and RF Diffusion solutions tended to be more similar than those which do not share similar solutions upstream of MPNN. Alignment and identity table by Clustal Omega. FIG. 7C shows the identity matrix for 16 selected J peptides.

[0012] FIG. 8 shows the iodixanol gradient purification of AAV9 produced by triple transfection method (XX680, pLH9, EFla EGFP J peptide), which demonstrated co-migration of EGFP deep into the gradient. Fractions 6 and 7 contained full particles packaging J peptide-EGFP fusions. Virus was harvested after 3 days from combined media / cells and freeze thawed to lyse. The cell pellet was clarified by centrifugation. The iodixanol gradient was prepared as in Zolotukhin et al., 1999.

[0013] FIG. 9A shows a graphical depiction of pLH9, CMV-AAV9 VP1, and CMV-AAV9 VP3 vector production plasmids showing deletions able to generate packaging vectors. FIG. 9B shows two example plasmids generated to test NLS-Cre or EGFP fusion to the J-peptide.

[0014] FIG. 10A shows that virus was produced by triple plasmid method for pLH9 / VP10RF, XX680 / pUC19, and EFla NLS-Cre. Cells were harvested at 3 days to examine expression of different combination of these plasmids. FIG. 10B shows that in the absence of Rep, the J peptide fusions still expressed. However, XX680 (pHelper) was required for robust expression of J peptides with capsid protein.

[0015] FIG. HA shows that vectors analyzed by mass photometry appeared to lack genomes as Rep and ITR were omitted but are otherwise indistinguishable from empty AAV9. Cre signal demonstrated loading of NLS-Cre. Titer estimates were determined by Refyen analysis. Vector produced by triple transfection method with XX680, pLH9, and NLS-Cre J peptide fusion cassette. FIG. 11B shows that virus was harvested after 3 days from combined media / cells and freeze thawed to lyse. Then, the cell pellet was clarified by centrifugation.

[0016] FIG. 12A - FIG. 12B show iodixanol gradient purification of AAV9 produced by triple transfection method (i.e., XX680 (pHelper), CMV-VP3 (trans-complementing plasmid - FIG. 12A), and EFla-Cre-J peptide (cis plasmid)). Fractions 5 through 7 contained full particles packaging J peptide-Cre fusions (FIG. 12B). Particles were harvested after 3 days from combined media / cells and freeze thawed to lyse. Then, the cell pellet was clarified by centrifugation.

[0017] FIG. 13A - FIG. 13B show iodixanol gradient purification of AAV9 produced by triple transfection method (i.e., XX680 (pHelper), CMV-VP1 (trans-complementing plasmid - FIG. 13A), EFla-Cre-J peptide (cis plasmid)). FIG. 13B shows fractions 4 through 7 contained full particles packaging J-peptide-Cre fusions. Particles were harvested after 3 days from combined media / cells and freeze thawed to lyse. Then, the cell pellet was clarified by centrifugation.V. BRIEF SUMMARY

[0018] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid.

[0019] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0020] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid.

[0021] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0022] Disclosed herein is a cargo that is linked to or fused with a disclosed peptide or a disclosed J peptide.

[0023] Disclosed herein is an AAV particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is an AAV-like particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a plurality of AAV particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a plurality AAV-like particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is an AAV particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is an AAV-like particlecomprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a plurality of AAV particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a plurality AAV-like particle comprising a disclosed cargo fused to a disclosed J peptide.

[0024] Disclosed herein is a method of loading a disclosed cargo into an AAV particle, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0025] Disclosed herein is a method of loading a cargo into an AAV particle, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0026] Disclosed herein is a method of loading a cargo into an AAV particle, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0027] Disclosed herein is a method of loading a disclosed cargo into an AAV-like particle, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0028] Disclosed herein is a method of loading a cargo into an AAV-like particle, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0029] Disclosed herein is a method of loading a cargo into an AAV-like particle, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0030] Disclosed herein is a method of generating an AAV particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0031] Disclosed herein is a method of generating an AAV particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0032] Disclosed herein is a method of generating an AAV particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0033] Disclosed herein is a method of generating an AAV-like particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0034] Disclosed herein is a method of generating an AAV-like particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0035] Disclosed herein is a method of generating an AAV-like particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).VI. DETAILED DESCRIPTION

[0036] The present disclosure describes formulations, compounded compositions, kits, capsules, containers, and / or methods thereof. It is to be understood that the inventive aspects of which are not limited to specific synthetic methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, example methods and materials are now described.

[0037] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention.A. Definitions

[0038] Before the present compounds, compositions, articles, systems, devices, and / or methods are disclosed and described, it is to be understood that they are not limited to specific synthetic methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purposeof describing particular aspects only and is not intended to be limiting. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, example methods and materials are now described.

[0039] This disclosure describes inventive concepts with reference to specific examples. However, the intent is to cover all modifications, equivalents, and alternatives of the inventive concepts that are consistent with this disclosure.

[0040] As used in the specification and the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise.

[0041] The phrase “consisting essentially of’ limits the scope of a claim to the recited components in a composition or the recited steps in a method as well as those that do not materially affect the basic and novel characteristic or characteristics of the claimed composition or claimed method. The phrase “consisting of’ excludes any component, step, or element that is not recited in the claim. The phrase “comprising” is synonymous with “including”, “containing”, or “characterized by”, and is inclusive or open-ended. “Comprising” does not exclude additional, unrecited components or steps.

[0042] In an aspect, when referring to any numerical value, the term “about” means a value falling within a range that is ± 10% of the stated value.

[0043] Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, a further aspect includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms a further aspect. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as “about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0044] References in the specification and concluding claims to parts by weight of a particular element or component in a composition denotes the weight relationship between the element or component and any other elements or components in the composition or article for which a part by weight is expressed. Thus, in a compound containing 2 parts by weight component X and 5 parts by weight component Y, X and Y are present at a weight ratio of 2:5, and are present in such ratio regardless of whether additional components are contained in the compound.

[0045] In an aspect, the terms “optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. In an aspect, a disclosed method can optionally comprise one or more additional steps, such as, for example, repeating an administering step or altering an administering step.

[0046] In an aspect, “isolated” refers to a nucleic acid molecule or a nucleic acid sequence that has been substantially separated, produced apart from, or purified away from other biological components in the cell or tissue of an organism in which the component occurs, such as other cells, chromosomal and extrachromosomal DNA and RNA, and proteins. Nucleic acids and proteins that have been “isolated” include nucleic acids and proteins purified by standard purification methods. The term also embraces nucleic acids and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids and proteins.

[0047] In an aspect, the term “subject” refers to the target of administration, e.g, a human being. The term “subject” also includes domesticated animals (e.g, cats, dogs, etc.), livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), and laboratory animals (e.g., mouse, rabbit, rat, guinea pig, fruit fly, etc.). Thus, the subject of the herein disclosed methods can be a vertebrate, such as a mammal, a fish, a bird, a reptile, or an amphibian. Alternatively, the subj ect of the herein disclosed methods can be a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig, or rodent. The term does not denote a particular age or sex, and thus, adult and child subjects, as well as fetuses, whether male or female, are intended to be covered. In an aspect, a subject can be a human patient. In an aspect, a subject can have a disease or disorder, be suspected of having a disease or disorder, or be at risk of developing a disease or disorder (e.g., a genetic disease or disorder).

[0048] In an aspect, a “regulatory element” can refer to promoters, enhancers, internal ribosomal entry sites (IRES), and other expression control elements (e.g., transcription termination signals, such as polyadenylation signals and poly-U sequences). Regulatory elements can include those that direct constitutive expression of a nucleotide sequence in many types of host cells and those that direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences).

[0049] In an aspect, the term “diagnosed” means having been subjected to an examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof, or by one or more of the disclosed methods. For example, “diagnosed with a disease or disorder” means having been subjected to an examination by a person of skill, forexample, a physician, and found to have a condition (such as a genetic disease or disorder) that can be treated by one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof, or by one or more of the disclosed methods. For example, “suspected of having a disease or disorder” can mean having been subjected to an examination by a person of skill, for example, a physician, and found to have a condition (such as a genetic disease or disorder) that can likely be treated by one or more of by one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof, or by one or more of the disclosed methods. In an aspect, an examination can be physical, can involve various tests (e.g., blood tests, genotyping, biopsies, etc.) and assays (e.g., enzymatic assay), or a combination thereof.

[0050] A “patient” refers to a subject afflicted with a disease or disorder (e.g., a genetic disease or disorder). In an aspect, a patient can refer to a subject that has been diagnosed with or is suspected of having a disease or disorder. In an aspect, a patient can refer to a subject that has been diagnosed with or is suspected of having a disease or disorder and is seeking treatment or receiving treatment for a disease or disorder.

[0051] In an aspect, the phrase “identified to be in need of treatment for a disease or disorder,” or the like, refers to selection of a subject based upon need for treatment of the disease or disorder. For example, a subject can be identified as having a need for treatment of a disease or disorder (e.g., a genetic disease or disorder) based upon an earlier diagnosis by a person of skill and thereafter subjected to treatment for the genetic disease or disorder. In an aspect, the identification can be performed by a person different from the person making the diagnosis. In an aspect, the administration can be performed by one who performed the diagnosis.

[0052] In an aspect, “inhibit,” “inhibiting”, and “inhibition” mean to diminish or decrease an activity, level, response, condition, severity, disease, or other biological parameter. This can include, but is not limited to, the complete ablation of the activity, level, response, condition, severity, disease, or other biological parameter. This can also include, for example, a 10% inhibition or reduction in the activity, level, response, condition, severity, disease, or other biological parameter as compared to the native or control level (e.g., a subject not having a disease or disorder such as a genetic disease or disorder). Thus, in an aspect, the inhibition or reduction can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of reduction in between as compared to native or control levels. In an aspect, the inhibition or reduction can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% as compared to native or control levels. In an aspect, the inhibition or reduction can be 0-25%, 25-50%, 50-75%, or 75-100% as compared to native or control levels. In an aspect, a native or control level can be a pre-disease or pre-disorder level.

[0053] The words “treat” or “treating” or “treatment” include palliative treatment, that is, treatment designed for the relief of symptoms rather than the curing of the disease, pathological condition, or disorder; preventative treatment, that is, treatment directed to minimizing or partially or completely inhibiting the development of the associated disease, pathological condition, or disorder; and supportive treatment, that is, treatment employed to supplement another specific therapy directed toward the improvement of the associated disease, pathological condition, or disorder. In an aspect, the terms cover any treatment of a subject, including a mammal (e.g., a human), and includes: (i) preventing the undesired physiological change, disease, pathological condition, or disorder from occurring in a subject that can be predisposed to the disease but has not yet been diagnosed as having it; (ii) inhibiting the physiological change, disease, pathological condition, or disorder, z.e., arresting its development; or (iii) relieving the physiological change, disease, pathological condition, or disorder, z.e., causing regression of the disease. For example, in an aspect, treating a disease or disorder can reduce the severity of an established a disease or disorder in a subject by 1%- 100% as compared to a control (such as, for example, an individual not having a genetic disease or disorder). In an aspect, treating can refer to a 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% reduction in the severity of a disease or disorder (such as a genetic disease or disorder). For example, treating a disease or disorder can reduce one or more symptoms of a disease or disorder in a subject by l%-100% as compared to a control (such as, for example, an individual not having a genetic disease or disorder). In an aspect, treating can refer to 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% reduction of one or more symptoms of an established a disease or disorder. It is understood that treatment does not necessarily refer to a cure or complete ablation or eradication of a disease or disorder. However, in an aspect, treatment can refer to a cure or complete ablation or eradication of a disease or disorder.

[0054] In an aspect, the term “prevent” or “preventing” or “prevention” refers to precluding, averting, obviating, forestalling, stopping, or hindering something from happening, especially by advance action. It is understood that where reduce, inhibit, or prevent are used herein, unless specifically indicated otherwise, the use of the other two words is also expressly disclosed. In an aspect, preventing a disease or disorder having chromatin deregulation and / or chromatin dysregulation is intended. The words “prevent”, “preventing”, and “prevention” also refer to prophylactic or preventative measures for protecting or precluding a subject (e.g., an individual) not having a given a disease or disorder (such as a genetic disease or disorder) a or related complication from progressing to that complication.

[0055] In an aspect, the “GH loop” refers to a loop sequence that is flanked by P-strand G and P- strand H within the internal P-barrel of the capsid protein. The “GH loop” sequence comprises variable region VR IV through VR VIII, including the encompassed GBS sequence and all interspersed conserved backbone sequence from the donor.

[0056] In an aspect, the terms “administering” and “administration” refer to any method of providing one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof to a subject. Such methods are well known to those skilled in the art and include, but are not limited to, the following: oral administration, transdermal administration, administration by inhalation, nasal administration, topical administration, in utero administration, intrahepatic administration, intravaginal administration, ophthalmic administration, intraaural administration, otic administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injectable such as intravenous administration, intra-CSF administration, intra-arterial administration, intramuscular administration, and subcutaneous administration. Administration can also include hepatic intraarterial administration or administration through the hepatic portal vein (HPV). Administration of a disclosed AAV particle or a disclosed AAV-like particle, a disclosed cargo, a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical composition, a disclosed therapeutic agent, a disclosed immune modulator, a disclosed proteasome inhibitor, a disclosed small molecule, a disclosed endonuclease, a disclosed oligonucleotide, and / or a disclosed RNA therapeutic can comprise administration directly into the CNS or the PNS. Administration can be continuous or intermittent. Administration can comprise a combination of one or more route.

[0057] In an aspect, the skilled person can determine an efficacious dose, an efficacious schedule, and an efficacious route of administration for one or more of the disclosed AAV particles, AAV- like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof to treat or prevent a disease or disorder (such as genetic disease or disorder). In an aspect, the skilled person can also alter, change, or modify an aspect of an administering step to improve efficacy of one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof.

[0058] By “determining the amount” is meant both an absolute quantification of a particular analyte (e.g., an mRNA sequence containing a particular tag) or a determination of the relative abundance of a particular analyte (e.g., an amount as compared to a mRNA sequence including a different tag). The phrase includes both direct or indirect measurements of abundance (e.g., individual mRNA transcripts may be quantified or the amount of amplification of an mRNAsequence under certain conditions for a certain period may be used a surrogate for individual transcript quantification) or both.

[0059] In an aspect, “modifying the method” can comprise modifying or changing one or more features or aspects of one or more steps of a disclosed method. For example, in an aspect, a method can be altered by changing the amount of one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof administered to a subject, or by changing the frequency of administration of one or more of the disclosed AAV particles, AAV- like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof to a subject, by changing the duration of time one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof are administered to a subject, or by substituting for one or more of the disclosed components and / or reagents with a similar or equivalent component and / or reagent. The same applies to all disclosed therapeutic agents, immune modulators, immunosuppressive agents, proteosome inhibitors, etc.

[0060] In an aspect, the term “pharmaceutically acceptable carrier” refers to sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents, or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. In an aspect, a pharmaceutical carrier employed can be a solid, liquid, or gas. In an aspect, examples of solid carriers can include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. In an aspect, examples of liquid carriers can include sugar syrup, peanut oil, olive oil, and water. In an aspect, examples of gaseous carriers can include carbon dioxide and nitrogen. In preparing a disclosed composition for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques. Proper fluidity can be maintained, for example, bythe use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid and the like. It can also be desirable to include isotonic agents such as sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents, such as aluminum monostearate and gelatin, which delay absorption. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, poly(orthoesters) and poly(anhydrides). Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues. The injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable media just prior to use. Suitable inert carriers can include sugars such as lactose. Desirably, at least 95% by weight of the particles of the active ingredient have an effective particle size in the range of 0.01 to 10 micrometers.

[0061] In an aspect, the term “excipient” refers to an inert substance which is commonly used as a diluent, vehicle, preservative, binder, or stabilizing agent, and includes, but is not limited to, proteins (e.g., serum albumin, etc.), amino acids (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylate, etc.), surfactants (e.g., SDS, polysorbate, nonionic surfactant, etc.), saccharides (e.g., sucrose, maltose, trehalose, etc.) and polyols (e.g., mannitol, sorbitol, etc.). See, also, for reference, Remington’s Pharmaceutical Sciences, (1990) Mack Publishing Co., Easton, Pa., which is hereby incorporated by reference in its entirety.

[0062] In an aspect, “concurrently” means (1) simultaneously in time, or (2) at different times during the course of a common treatment schedule.

[0063] In an aspect, the term “contacting” refers to bringing one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof together with a target area or intended target area in such a manner that the one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof exert an effect on the intended target or targeted area either directly or indirectly. For example, a disclosed J peptide can becontacted with a disclosed cargo and a disclosed fusion between a J peptide and a cargo can be contacted with the luminal surface of an AAV particle or AAV-like particle. A target area can comprise one or more cells, and in an aspect, one or more cells can be in a subject. A target area or intended target area can be one or more of a subject’s organs (e.g., lungs, heart, liver, kidney, brain, etc.). In an aspect, a target area or intended target area can be any cell or any organ infected by a disease or disorder (such as a genetic disease or disorder). In an aspect, a target area or intended target area can be any organ, tissue, or cells that are affected by a disease or disorder (such as a genetic disease or disorder).

[0064] In an aspect, “determining” can refer to measuring or ascertaining the presence and severity of a disease or disorder, such as, for example, a genetic disease or disorder. Methods and techniques used to determine the presence and / or severity of a disease or disorder are typically known to the medical arts. For example, the art is familiar with the ways to identify and / or diagnose the presence, severity, or both of a disease or disorder (such as, for example, a genetic disease or disorder).

[0065] In an aspect, “effective amount” and “amount effective” can refer to an amount that is sufficient to achieve the desired result such as, for example, the treatment and / or prevention of a disease or disorder (e.g., a genetic disease or disorder) or a suspected disease or disorder. In an aspect, the terms “effective amount” and “amount effective” can refer to an amount that is sufficient to achieve the desired an effect on an undesired condition e.g., a disease or disorder).

[0066] For example, a “therapeutically effective amount” refers to an amount that is sufficient to achieve the desired therapeutic result or to have an effect on undesired symptoms, but is generally insufficient to cause adverse side effects. In an aspect, “therapeutically effective amount” means an amount of a disclosed AAV particle or a disclosed AAV-like particle, a disclosed cargo, a disclosed nucleic acid molecule, a disclosed vector, or a disclosed pharmaceutical formulation; that (i) treats the particular disease, condition, or disorder (e.g., a genetic disease or disorder), (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder e.g., a genetic disease or disorder), or (iii) delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein (e.g., a genetic disease or disorder). The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the disclosed AAV particles or disclosed AAV-like particles, the disclosed cargos, the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations employed; the disclosed methods employed; the age, body weight, general health, sex and diet of the patient; the time of administration; the route of administration; the rate of excretion of the disclosed AAV particles or disclosed AAV-like particles, the disclosed cargo, the disclosed nucleic acidmolecules, disclosed vectors, or disclosed pharmaceutical formulations employed; the duration of the treatment; drugs used in combination or coincidental with the disclosed AAV particles or disclosed AAV-like particles, the disclosed cargo, the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations employed, and other like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, then the effective daily dose can be divided into multiple doses for purposes of administration. Consequently, a single dose of the disclosed AAV particles or disclosed AAV-like particles, the disclosed cargo, the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations can contain such amounts or submultiples thereof to make up the daily dose. The dosage can be adjusted by the individual physician in the event of any contraindications. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products. In further various aspects, a preparation can be administered in a “prophylactically effective amount”; that is, an amount effective for prevention of a disease or condition, such as, for example, a disease or disorder due to a missing, deficient, and / or mutant protein or enzyme.

[0067] In an aspect, “operably linked” means that expression of a gene or a transgene is under the control of a promoter with which it is spatially connected. A promoter can be positioned 5’ (upstream) or 3’ (downstream) of a gene under its control. The distance between the promoter and a gene can be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. As is known in the art, variation in this distance can be accommodated without loss of promoter function.

[0068] In an aspect, “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein must contain at least two amino acids and there is no limitation on the maximum number of amino acids that can comprise a protein’s sequence. The term “peptide” can refer to a short chain of amino acids including, for example, natural peptides, recombinant peptides, synthetic peptides, or any combination thereof. Proteins and peptides can include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, and fusion proteins, among others.

[0069] “Nucleic acid” or “oligonucleotide” or “polynucleotide” In an aspect means at least two nucleotides covalently linked together. The depiction of a single strand can also define thesequence of the complementary strand. Thus, a nucleic acid can encompass the complementary strand of a depicted single strand. Many variants of a nucleic acid can be used for the same purpose as a given nucleic acid. Thus, a nucleic acid can encompass substantially identical nucleic acids and complements thereof. A single strand can provide a probe that can hybridize to a target sequence under stringent hybridization conditions. Thus, a nucleic acid can encompass a probe that hybridizes under stringent hybridization conditions. A nucleic acid can be single-stranded, or double-stranded, or can contain portions of both double-stranded and single-stranded sequence. The nucleic acid can be DNA, both genomic and cDNA, RNA, or a hybrid, where the nucleic acid can contain combinations of deoxyribo- and ribo-nucleotides, and combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine. Nucleic acids can be obtained by chemical synthesis methods or by recombinant methods. Also, in an aspect, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid construct,” “nucleotide sequence”, and “polynucleotide” can refer to RNA or DNA that is linear or branched, single or double stranded, or a hybrid thereof. The term can encompass RNA / DNA hybrids. When dsRNA is produced synthetically, less common bases, such as inosine, 5- methylcytosine, 6-methyladenine, hypoxanthine and others can also be used for antisense, dsRNA, and ribozyme pairing. For example, polynucleotides that contain C-5 propyne analogues of uridine and cytidine have been shown to bind RNA with high affinity and to be potent antisense inhibitors of gene expression. Other modifications, such as modification to the phosphodiester backbone, or the 2’-hydroxy in the ribose sugar group of the RNA can also be made. A “synthetic” nucleic acid or polynucleotide, in an aspect, refers to a nucleic acid or polynucleotide that is not found in nature but is constructed by the hand of man and therefore is not a product of nature.

[0070] A “polynucleotide” is a sequence of nucleotide bases, and may be RNA, DNA, or DNA- RNA hybrid sequences (including both naturally occurring and non-naturally occurring nucleotides).

[0071] A “fragment” or “portion” of a nucleotide sequence can be understood to mean a nucleotide sequence of reduced length relative (e.g., reduced by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more nucleotides) to a reference nucleic acid or nucleotide sequence and comprising, consisting essentially of, or consisting of a nucleotide sequence of contiguous nucleotides identical or almost identical (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical) to the reference nucleic acid or nucleotide sequence. Such a nucleic acid fragment or portion according to the disclosure can be, where appropriate, included in a larger polynucleotide of which it is a constituent. In an aspect, a fragment or portionof a nucleotide sequence or nucleic acid sequence can comprise the sequence encoding an exon having one or more mutations.

[0072] A “fragment” or “portion” of an amino acid sequence can be understood to mean an amino acid sequence of reduced length relative (e.g., reduced by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, or more amino acids) to a reference amino acid sequence and comprising, consisting essentially of, or consisting of an amino acid sequence of contiguous amino acids identical or almost identical (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical) to the reference amino acid sequence. Such an amino acid fragment or portion according to the disclosure can be, where appropriate, included in a larger amino acid sequence of which it is a constituent.

[0073] A “heterologous” or a “recombinant” nucleotide or amino acid sequence as used interchangeably herein can refer to a nucleotide or an amino acid sequence not naturally associated with a host cell into which it is introduced, including non-naturally occurring multiple copies of a naturally occurring nucleotide or amino acid sequence.

[0074] Different nucleic acids or proteins having homology can be referred to as “homologues”. The term homologue includes homologous sequences from the same and other species and orthologous sequences from the same and other species. “Homology” refers to the level of similarity between two or more nucleic acid and / or amino acid sequences in terms of percent of positional identity (i.e., sequence similarity or identity). Homology also refers to the concept of similar functional properties among different nucleic acids or proteins. Thus, the disclosed compositions and disclosed methods can comprise homologues to the disclosed nucleotide sequences and / or disclosed polypeptide sequences.

[0075] In an aspect, “orthologous” can refer to homologous nucleotide sequences and / or amino acid sequences in different species that arose from a common ancestral gene during speciation. A homologue of a disclosed nucleotide sequence or a disclosed polypeptide can have substantial sequence identity (e.g., at least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, and / or 100%) to a disclosed nucleotide sequence or a disclosed polypeptide.

[0076] In an aspect, “Complement” or “complementary” means a nucleic acid can mean Watson- Crick (e.g., A-T / U and C-G) or Hoogsteen base pairing between nucleotides or nucleotide analogs of nucleic acid molecules. “Complementarity” refers to a property shared between two nucleic acid sequences, such that when they are aligned antiparallel to each other, the nucleotide bases at each position will be complementary.

[0077] In an aspect, “promoter” or “promoters” are known to the art. Depending on the level and tissue-specific expression desired, a variety of promoter elements can be used. A promoter can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the gene expression desired. A promoter can be native (endogenous) or foreign (exogenous) and can be a natural or a synthetic sequence. By foreign or exogenous, it is intended that the transcriptional initiation region is not found in the wild-type host into which the transcriptional initiation region is introduced.

[0078] “Tissue-specific promoters” are known to the art and include, but are not limited to, neuron-specific promoters, muscle-specific promoters, liver-specific promoters, skeletal musclespecific promoters, and heart-specific promoters.

[0079] “Liver-specific promoters” are known to the art and include, but are not limited to, the thyroxin binding globulin (TBG) promoter, the al-microglobulin / bikunin enhancer / thyroid hormone-binding globulin promoter, the human albumin (hALB) promoter, the thyroid hormone- binding globulin promoter, the a- 1 -anti -trypsin promoter, the bovine albumin (bAlb) promoter, the murine albumin (mAlb) promoter, the human al -antitrypsin (hAAT) promoter, the ApoEhAAT promoter comprising the ApoE enhancer and the hAAT promoter, the transthyretin (TTR) promoter, the liver fatty acid binding protein promoter, the hepatitis B virus (HBV) promoter, the DC 172 promoter comprising the hAAT promoter and the al -microglobulin enhancer, the DC 190 promoter comprising the human albumin promoter and the prothrombin enhancer, or any other natural or synthetic liver-specific promoter. In an aspect, a liver specific promoter can comprise about 845-bp and comprise the thyroid hormone-binding globulin promoter sequences (2382 to 13), two copies of al-microglobulin / bikunin enhancer sequences (22,804 through 22,704), and a 71-bp leader sequence as described by Ill CR, et al. (1997). In an aspect, a disclosed liver specific promoter can comprise the sequence set forth in SEQ ID NO:32, or a sequence having about 50%, about 60%, about 70% about 80%, about 90%, about 95%, or more identity to the sequence set forth in SEQ ID NO:32.

[0080] Ubiquitous / constitutive promoters” are known to the art and include, but are not limited to, a CMV major immediate-early enhancer / chicken beta-actin promoter, a cytomegalovirus (CMV) major immediate-early promoter, an Elongation Factor 1-a (EFl -a) promoter, a simian vacuolating virus 40 (SV40) promoter, an AmpR promoter, a PyK promoter, a human ubiquitin C gene (Ubc) promoter, a MFG promoter, a human beta actin promoter, a CAG promoter, a EGR1 promoter, a FerH promoter, a FerL promoter, a GRP78 promoter, a GRP94 promoter, a HSP70 promoter, a Q-kin promoter, a murine phosphoglycerate kinase (mPGK) or human PGK (hPGK) promoter, a ROSA promoter, human Ubiquitin B promoter, a Rous sarcoma virus promoter, or any other natural or synthetic ubiquitous / constitutive promoters.

[0081] In an aspect, an “inducible promoter” refers to a promoter that can be regulated by positive or negative control. Factors that can regulate an inducible promoter include, but are not limited to, chemical agents (e.g., the metallothionein promoter or a hormone inducible promoter), temperature, and light.

[0082] In an aspect, the term “serotype” is a distinction used to refer to an AAV having a capsid that is serologically distinct from other AAV serotypes. Serologic distinctiveness can be determined by the lack of cross-reactivity between antibodies to one AAV as compared to another AAV. Such cross-reactivity differences are usually due to differences in capsid protein sequences / antigenic determinants (e.g., due to VP1, VP2, and / or VP3 sequence differences of AAV serotypes).

[0083] In an aspect, “tropism” refers to the specificity of an AAV capsid protein present in an AAV viral particle, for infecting a particular type of cell or tissue. The tropism of an AAV capsid for a particular type of cell or tissue may be determined by measuring the ability of AAV vector particles comprising the hybrid AAV capsid protein to infect or to transduce a particular type of cell or tissue, using standard assays that are well-known in the art such as those disclosed in the examples of the present application. In an aspect, the term “liver tropism” or “hepatic tropism” refers to the tropism for liver or hepatic tissue and cells, including hepatocytes.

[0084] “Sequence identity” and “sequence similarity” can be determined by alignment of two peptide or two nucleotide sequences using global or local alignment algorithms. Sequences may then be referred to as “substantially identical” or “essentially similar” when they are optimally aligned. For example, sequence similarity or identity can be determined by searching against databases such as FASTA, BLAST, etc., but hits should be retrieved and aligned pairwise to compare sequence identity. Two proteins or two protein domains, or two nucleic acid sequences can have “substantial sequence identity” if the percentage sequence identity is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99% or more, preferably 90%, 95%, 98%, 99% or more. Such sequences are also referred to as “variants” herein, e.g., other variants of a missing, deficient, and / or mutant protein or enzyme. It should be understood that sequence with substantial sequence identity do not necessarily have the same length and may differ in length. For example, sequences that have the same nucleotide sequence but of which one has additional nucleotides on the 3’- and / or 5’-side are 100% identical.

[0085] In an aspect, “codon optimization” can refer to a process of modifying a nucleic acid sequence for enhanced expression in the host cells of interest by replacing one or more codons or more of the native sequence with codons that are more frequently or most frequently used in the genes of that host cell while maintaining the native amino acid sequence. Various species exhibit particular bias for certain codons of a particular amino acid. As contemplated herein, genes canbe tailored for optimal gene expression in a given organism based on codon optimization. Codon usage tables are readily available, for example, at the “Codon Usage Database.” Many methods and software tools for codon optimization have been reported previously. (See, for example, genomes.urv.es / OPTIMIZER / ).

[0086] In an aspect, “RNA Editing” can refer to a type of genetic engineering in which an RNA molecule (or ribonucleotides of the RNA) is inserted, deleted, or replaced in the genome of an organism using engineered nucleases (such as the Casl3d proteins provided herein), which create site-specific strand breaks at desired locations in the RNA. The induced breaks are repaired resulting in targeted mutations or repairs.

[0087] In an aspect, “CRISPR or clustered regularly interspaced short palindromic repeat” is an ideal tool for correction of genetic abnormalities as the system can be designed to target genomic DNA directly. Cas9 is well-known to the art. The CRISPR / Cas methods disclosed herein, such as those that use an Cast 3d, can be used to edit the sequence of one or more target RNAs, such as one associated with a disease or disorder disclosed herein (e.g., a genetic disease or disorder).

[0088] The diverse Casl3 family contains at least four known subtypes, including Casl3a (formerly C2c2), Cast 3b, Cast 3 c, and Cast 3d. All known Cast 3 family members contain two HEPN domains, which confer RNase activity. Casl3 can be reprogrammed to cleave a targeted ssRNA molecule through a short guide RNA with complementarity to the target sequence. Cas 13s function similarly to Cas9, using a ~64-nucleotide guide RNA to encode target specificity. The Cas 13 protein complexes with the guide RNA via recognition of a short hairpin in the crRNA, and target specificity is encoded by a 28-nucleotide to a 30-nucleotide spacer that is complementary to the target region. In addition to programmable RNase activity, all Casl3s exhibit collateral activity after recognition and cleavage of a target transcript, leading to nonspecific degradation of any nearby transcripts regardless of complementarity to the spacer. While Cas 13a showed some activity for RNA knockdown, certain orthologs of Cas 13b proved more stable and robust in mammalian cells for RNA knockdown and editing. More recently, additional orthologs of Cas 13 have been discovered, including Cas 13d, which has been leveraged for efficient and robust knockdown across many endogenous transcripts. Casl3d can be used to modulate splicing of endogenous transcripts and that the coding sequence for Casl3d is small enough to fit within the packaging limits of AAV for in vivo delivery.

[0089] In an aspect, Cas 13 can be considered an outlier in the CRISPR world because it targets RNA, not DNA. Once it is activated by a ssRNA sequence bearing complementarity to its crRNA spacer, it unleashes a nonspecific RNase activity and destroys all nearby RNA regardless of their sequence. As disclosed herein, this property can be harnessed in vitro for precision diagnostics. Generally, Casl3 can be found in Leptotrichia buccalis, Leptotrichia shahii, Ruminococcusflavefaciens, Bergeyella zoohelcum, Prevotella buccae, and Listeria seeligeri and can have a size of about 900 to about 1300 amino acids. In an aspect, the guide spacer length can be about 22 to about 30 nucleotides while the total guide length can be about 52 to about 66 nucleotides. In an aspect, a PAM can be 3-H for LshCasl3a, 5-D and 3-NAN or NNA for BzCasl3b, and none for RfCasl3d. In an aspect, a disclosed Casl3 can cut ssRNA.

[0090] As known to the skilled person in the cart, a Cast 3d ortholog can be from a prokaryotic genome or metagenome, gut metagenome, an activated sludge metagenome, an anaerobic digester metagenome, a chicken gut metagenome, a human gut metagenome, a pig gut metagenome, a bovine gut metagenome, a sheep gut metagenome, a goat gut metagenome, a capybara gut metagenome, a primate gut metagenome, a termite gut metagenome, a fecal metagenome, a genome from the Order Clostridiales, or the Family Ruminococcaceae. In an aspect, a disclosed Cast 3d ortholog can include an Cast 3d ortholog from Ruminococcus albus, Eubacterium siraeum, a Ruminococcus flavefaciens strain XPD3002, Ruminococcus flavefaciens FD-1, uncultured Eubacterium sp TS28-c4095, uncultured Ruminococcus sp., Ruminococcus bicirculans, or Ruminococcus sp CAG57.

[0091] As known to the art, RNA binding proteins consist of multiple repetitive sequences that contain only a few specific basic domains. Structurally, common RNA-binding domains mainly include RNA-recognition motif (RRM), K homology (KH) domain, double-stranded RBD (dsRBD), cold-shock domain (CSD), arginine-glycine-glycine (RGG) motif, tyrosine-rich domain, and zinc fingers (ZnF) of the CCHC, CCCH, ZZ type etc. According to the different functions of RBPs in cells, RBPs can be divided into epithelial splicing regulatory proteins (ESRP1), cytoplasmic polyadenylation element binding protein family (CPEB1 / 2), Hu-antigen R (HuR), heterogeneous nuclear ribonucleoprotein family members (hnRNP A / D / H / K / M / E / L), insulin-like growth factor 2 mRNA family members (IMP 1 / 2 / 3), zfh family of transcription factors (ZEB 1 / 2), KH-type splicing regulatory protein (KHSRP), La ribonucleoprotein domain family members (LARP 1 / 6 / 7), Lin-28 homolog proteins (Lin28), Musashi protein family (MSI1 / 2), Pumilio protein family (PUM1 / 2), Quaking (QK), RNA-binding motif protein family (4 / 10 / 38 / 47), Src-associated substrate during mitosis of 68 kDa (SAM68), serine and arginine rich splicing factor (SRSF1 / 3), T cell intracellular antigens (TIA1 / TIAR), and Upstream of N-Ras (UNR).

[0092] In an aspect, the term “in silico” can mean experimentation performed by computer. In an aspect, in silico experimentation can be performed first and then in silico results can be validated in a series of in vitro and in vivo examples (as described herein in the Examples).

[0093] In an aspect, “immune tolerance,” “immunological tolerance,” and “immunotolerance” refers to a state of unresponsiveness or blunted response of the immune system to substances (e.g.,the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof, etc.) that have the capacity to elicit an immune response in a subject. Immune tolerance is induced by prior exposure to a specific antigen. Immune tolerance can be determined in a subject by measuring antibodies against a particular antigen or by liver-restricted transgene expression with a viral vector (such as, for example, AAV). Low or absent antibody titers over time is an indicator of immune tolerance. For example, in some embodiments, immune tolerance can be established by having IgG antibody titers of less than or equal to about 12,000, 11,500, 11,000, 10,500, 10,000, 9,500, 9,000, 8,500, 8,000, 7,500, 7,000, 6,500, or 6,000 within following gene therapy (such as the administration of the transgene encoding, for example, a missing, deficient, and / or mutant protein or enzyme).

[0094] As known to the art, antibodies (Abs) can mitigate AAV infection through multiple mechanisms by binding to AAV capsids and blocking critical steps in transduction such as cell surface attachment and uptake, endosomal escape, productive trafficking to the nucleus, or uncoating as well as promoting AAV opsonization by phagocytic cells, thereby mediating their rapid clearance from the circulation. For example, in humans, serological studies reveal a high prevalence of NAbs in the worldwide population, with about 67% of people having antibodies against AAV1, 72% against AAV2, and approximately 40% against AAV serotypes 5 through 9. Vector immunogenicity represents a major challenge in re-administration of AAV vectors.

[0001] In an aspect, also disclosed herein are partial self-complementary parvovirus (e.g., a disclosed AAV) genomes, plasmid vectors encoding the parvovirus genomes, and parvovirus (e.g., a disclosed AAV) particles including such genomes. In an aspect, provided herein is a plasmid vector comprising a nucleotide sequence encoding a disclosed parvovirus genome such as for example, a disclosed AAV. In an aspect, provided herein is a partial self-complementary parvovirus genome including a payload construct, parvovirus ITRs flanking the payload construct, and a self-complementary region flanking one of the ITRs. A self-complementary region can comprise a nucleotide sequence that is complementary to the payload construct. A disclosed self- complementary region can have a length that is less the entire length of the payload construct.

[0002] In an aspect, a disclosed self-complementary region of a disclosed parvovirus genome can comprise a minimum length, while still having a length that is less the entire length of the payload construct. In an aspect, a disclosed self-complementary region can comprise at least 50 bases in length, at least 100 bases in length, at least 200 in length, at least 300 bases in length, at least 400 bases in length, at least 500 bases in length, at least 600 bases in length, at least 700 bases in length, at least 800 bases in length, at least 900 bases in length, or at least 1,000 bases in length.

[0003] In an aspect, a “self-complementary parvovirus genome” can be a single stranded polynucleotide having, in the 5’ to 3’ direction, a first parvovirus ITR sequence, a heterologous sequence (e.g., payload construct comprising, for example, a desired gene), a second parvovirus ITR sequence, a second heterologous sequence, wherein the second heterologous sequence is complementary to the first heterologous sequence, and a third parvovirus ITR sequence. In contrast to a self-complementary genome, a “partial self-complementary genome” does not include three parvovirus ITRs and the second heterologous sequence that is complementary to the first heterologous sequence has a length that is less than the entire length of the first heterologous sequence (e.g., payload construct). Accordingly, a partial self-complementary genome is a single stranded polynucleotide having, in the 5’ to 3’ direction or the 3’ to 5’ direction, a first parvovirus ITR sequence, a heterologous sequence (e.g., payload construct), a second parvovirus ITR sequence, and a self-complementary region that is complementary to a portion of the heterologous sequence and has a length that is less than the entire length the heterologous sequence.

[0095] In an aspect, “immune-modulating” refers to the ability of one or more of the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof to alter (modulate) one or more aspects of the immune system. The immune system functions to protect the organism from infection and from foreign antigens by cellular and humoral mechanisms involving lymphocytes, macrophages, and other antigen-presenting cells that regulate each other by means of multiple cell-cell interactions and by elaborating soluble factors, including lymphokines and antibodies, that have autocrine, paracrine, and endocrine effects on immune cells.

[0096] In an aspect, “immune modulator” refers to an agent that is capable of adjusting a given immune response to a desired level (e.g., as in immunopotentiation, immunosuppression, or induction of immunologic tolerance). Examples of immune modulators include but are not limited to, a disclosed immune modulator can comprise aspirin, azathioprine, belimumab, betamethasone dipropionate, betamethasone valerate, bortezomib, bredinin, cyazathioprine, cyclophosphamide, cyclosporine, deoxyspergualin, didemnin B, fluocinolone acetonide, folinic acid, ibuprofen, IL6 inhibitors (such as sarilumab) indomethacin, inebilizumab, intravenous gamma globulin (IVIG), methotrexate, methylprednisolone, mycophenolate mofetil, naproxen, prednisolone, prednisone, prednisolone indomethacin, rapamycin, rituximab, sirolimus, sulindac, synthetic vaccine particles containing rapamycin (SVP -Rapamycin or ImmTOR), thalidomide, tocilizumab, tolmetin, triamcinolone acetonide, anti-CD3 antibodies, anti-CD4 antibodies, anti-CD19 antibodies, anti- CD20 antibodies, anti-CD22 antibodies, anti-CD40 antibodies, anti-FcRN antibodies, anti-IL6 antibodies, anti -IGF 1R antibodies, an IL2 mutein, a BTK inhibitor, or a combination thereof. In an aspect, a disclosed immune modulator can comprise one or more Treg (regulatory T cells)infusions (e.g., antigen specific Treg cells to AAV). In an aspect, a disclosed immune modulator can be bortezomib or SVP-Rapamycin. In an aspect, an immune modulator can be administered by any suitable route of administration including, but not limited to, in utero, intra-CSF, intrathecally, intravenously, subcutaneously, transdermally, intradermally, intramuscularly, orally, transcutaneously, intraperitoneally (IP), or intravaginally. In an aspect, a disclosed immune modulator can be administered using a combination of routes. Administration can also include hepatic intra-arterial administration or administration through the hepatic portal vein (HPV). Administration of an immune modulator can be continuous or intermittent, and administration can comprise a combination of one or more routes.

[0097] In an aspect, the term “immunotolerant” refers to unresponsiveness to an antigen (e.g., a vector, a therapeutic protein, a transgene product, etc.). An immunotolerant promoter can reduce, ameliorate, or prevent transgene-induced immune responses that can be associated with gene therapy. Assays known in the art to measure immune responses, such as immunohistochemical detection of cytotoxic T cell responses, can be used to determine whether one or more promoters can confer immunotolerant properties.

[0098] In an aspect, the term “package insert” is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications and / or warnings concerning the use of such therapeutic products.

[0099] In an aspect, the term “in combination” in the context of the administration of other therapies (e.g., other agents) includes the use of more than one therapy (e.g., drug therapy). Administration “in combination with” one or more further therapeutic agents includes simultaneous (e.g., concurrent) and consecutive administration in any order. The use of the term “in combination” does not restrict the order in which therapies are administered to a subject. By way of non-limiting example, a first therapy (e.g., the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof) may be administered prior to (e.g., 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks), concurrently, or after (e.g., 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks or longer) the administration of a second therapy (e.g., agent) to a subject having or diagnosed with a disease or disorder (such as a genetic disease or disorder).

[0100] Disclosed are the components to be used to prepare the disclosed AAV particles, AAV- like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof as well as the disclosed AAV particles, AAV-like particles, disclosed cargos, disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof used within the methods disclosed herein. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds cannot be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed and a number of modifications that can be made to a number of molecules including the compounds are discussed, specifically contemplated is each and every combination and permutation of the compound and the modifications that are possible unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E, and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited each is individually and collectively contemplated meaning combinations, A-E, A-F, B-D, B-E, B-F, C- D, C-E, and C-F are considered disclosed. Likewise, any subset or combination of these is also disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the compositions of the invention. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the methods of the invention.B. Compositions1. J Peptides

[0101] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid.

[0102] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the luminal surface of arecombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0103] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15.

[0104] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11.

[0105] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQID NO:11. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:11. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:11.

[0106] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and comprising the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0107] Disclosed herein is a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid and comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid and a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.Table 1 - Listing of Exemplary J Peptides

[0108] In an aspect, a disclosed peptide can be referred to as a “J peptide”. In an aspect, a disclosed peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid can be referred to as a “J peptide”. In an aspect, a disclosed peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid can be referred to as a “J peptide”. In an aspect, a disclosed peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid can be referred to as a “J peptide”. In an aspect, a disclosed peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid can be referred to as a “J peptide”.

[0109] In an aspect, a disclosed peptide can comprise 20 to 40 amino acids. In an aspect, a disclosed peptide can comprise 20 to 30 amino acids. In an aspect, a disclosed peptide can comprise 30 to 40 amino acids. In an aspect, a disclosed peptide can comprise 25 to 35 amino acids. In an aspect, a disclosed peptide can comprise 30 amino acids. In an aspect, a disclosed peptide can comprise 20 amino acids, 21 amino acids, 22 amino acids, 23 amino acids, 24 aminoacids, 25 amino acids, 26 amino acids, 27 amino acids, 28 amino acids, 29 amino acids, 30 amino acids, 31 amino acids, 32 amino acids, 33 amino acids, 34 amino acids, 35 amino acids, 36 amino acids, 37 amino acids, 38 amino acids, 39 amino acids, or 40 amino acids. In an aspect, a disclosed peptide can comprise at least 20 amino acids, at least 21 amino acids, at least 22 amino acids, at least 23 amino acids, at least 24 amino acids, at least 25 amino acids, at least 26 amino acids, at least 27 amino acids, at least 28 amino acids, at least 29 amino acids, at least 30 amino acids, at least 31 amino acids, at least 32 amino acids, at least 33 amino acids, at least 34 amino acids, 35 amino acids, at least 36 amino acids, 37 amino acids, at least 38 amino acids, at least 39 amino acids, or at least 40 amino acids.

[0110] In an aspect, a disclosed peptide can comprise the sequence of any one of SEQ ID NO:01 to SEQ ID NO:49. In an aspect, a disclosed peptide can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:49. In an aspect, a disclosed peptide can comprise a sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more than at least 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:49. In an aspect, a disclosed peptide can comprise a sequence having about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more than about 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:49. In an aspect, a disclosed peptide can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:49. In an aspect, a disclosed peptide can be any disclosed peptide but for one of SEQ ID NO:30 - SEQ ID NO:49.

[0111] In an aspect, a disclosed peptide can comprise the sequence of any one of SEQ ID NO:01 to SEQ ID NO:29. In an aspect, a disclosed peptide can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. In an aspect, a disclosed peptide can comprise a sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more than at least 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. In an aspect, a disclosed peptide can comprise a sequence having about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more than about 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. In an aspect, a disclosed peptide can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0112] In an aspect, a disclosed peptide can comprise the sequence of any one of SEQ ID NO:01 to SEQ ID NO: 15. In an aspect, a disclosed peptide can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. In an aspect, a disclosed peptide can comprise a sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more than at least 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. In an aspect, a disclosed peptide can comprise a sequence having about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more than about 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. In an aspect, a disclosed peptide can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0113] In an aspect, a disclosed peptide can comprise the sequence of any one of SEQ ID NO:01 to SEQ ID NO: 11. In an aspect, a disclosed peptide can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:11. In an aspect, a disclosed peptide can comprise a sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more than at least 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. In an aspect, a disclosed peptide can comprise a sequence having about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more than about 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:11. In an aspect, a disclosed peptide can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11.

[0114] In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:01 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:50. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:02 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:51. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:03 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:52. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:04 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:53. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:05 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:54. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:06 can be encoded by the nucleotide sequence comprisingthe sequence set forth in SEQ ID NO:55. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:07 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:56. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:08 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:57. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:09 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:58. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 10 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:59. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:11 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:60. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 12 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:61. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 13 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:62. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 14 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:63. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 15 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:64. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 16 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:65. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 17 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:66. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 18 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:67. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO: 19 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:68. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:20 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:69. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:21 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:70. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:22 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:71. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:23 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:72. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:24 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:73. In an aspect, a disclosed peptidecomprising the sequence set forth in SEQ ID NO:25 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:74. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:26 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:75. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:27 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:76. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:28 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:77. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:29 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:78. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:30 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:79. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:31 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:80. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:32 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:81. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:33 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:82. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:34 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:83. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:35 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:84. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:36 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:85. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:37 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:86. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:38 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:87. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:39 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:88. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:40 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:89. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:41 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:90. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:42 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:91. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:43 can be encoded by thenucleotide sequence comprising the sequence set forth in SEQ ID NO:92. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:44 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:93. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:45 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:94. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:46 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:95. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:47 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:96. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:48 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:97. In an aspect, a disclosed peptide comprising the sequence set forth in SEQ ID NO:49 can be encoded by the nucleotide sequence comprising the sequence set forth in SEQ ID NO:98.

[0115] In an aspect, a disclosed J peptide can be encoded by a sequence having about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more than about 95% identity to the sequence set forth in any one of SEQ ID NO:49 - SEQ ID NO:98. In an aspect, a disclosed J peptide can be encoded by a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:49 - SEQ ID NO:98.

[0116] In an aspect, a disclosed peptide can comprise a predicted aligned error (PAE) score of less than 10. In an aspect, a disclosed peptide can comprise a predicted aligned error (PAE) score of less than 7. In an aspect, PAE score is defined as the expected position error at residue X, measured in Angstroms, if the predicted and actual structures were aligned on residue Y. In an aspect, PAE score is measured using AlphaFold2. In an aspect, a low PAE score between two residues from two different domains can mean low predicted error. In an aspect, a low PAE score can mean that AlphaFold2 is confident in the position of these residues. In an aspect, a high PAE score can mean that AlphaFold2 is not confident in the relative positions of two residues.

[0117] In an aspect, a disclosed peptide or a disclosed J peptide can be fused to a cargo. In an aspect, a disclosed peptide or a disclosed J peptide fused to a cargo can comprise the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. In an aspect, a disclosed peptide or a disclosed J peptide fused to a cargo can comprising a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0118] In an aspect, a disclosed peptide or a disclosed J peptide fused to a cargo be used with AAV particles. In an aspect, a disclosed peptide or a disclosed J peptide fused to a cargo be usedwith AAV-like particles. In an aspect, AAV-like particles can be empty AAV capsids and / or can be devoid of any viral genomic material. In an aspect, a disclosed peptide or a disclosed J peptide fused to a cargo be used with AAV particles to create a nanocarrier. In an aspect, a disclosed peptide or a disclosed J peptide fused to a cargo be used with AAV-like particles to create a nanocarrier.

[0119] Disclosed herein is a library of J peptides. In an aspect, a disclosed library can comprise 2O30J peptides. In an aspect, a disclosed library can comprise J peptides having a PAE less than 10. In an aspect, a disclosed library can comprise J peptides having a PAE less than 7. Disclosed herein is a library of J peptides, each having the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:49. Disclosed herein is a library of J peptides, each having a sequence a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:49. Disclosed herein is a library of J peptides, each having the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. Disclosed herein is a library of J peptides, each having a sequence a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:11. Disclosed herein is a library of J peptides, each having the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15. Disclosed herein is a library of J peptides, each having a sequence a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15. Disclosed herein is a library of J peptides, each having the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

[0120] Disclosed herein is a library of J peptides, each having a sequence a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

[0121] Disclosed herein is a library of J peptides, each having a sequence a sequence having about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or more than about 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

[0122] In an aspect, a disclosed AAV capsid can comprise the capsid of AAV1, AAV2, AAV3 (including 3a and 3b), AAV4, AAV5, AAV6, AAV6.2, AAV7, AAV8, AAVrh8, AAV9, AAV10, AAVrhlO, AAV11, AAV12, or AAV13. In an aspect, a disclosed AAV capsid can comprise the capsid of a non-human species including but not limited to bovine AAV, caprine AAV, canine AAV, equine AAV, ovine AAV, avian AAV, non-human primate AAV, non-primate AAV, or reptilian AAV. In an aspect, a disclosed AAV capsid can comprise the capsid of any other virus classified by the International Committee on Taxonomy of Viruses (ICTV) as an AAV.

[0123] In an aspect, a disclosed AAV capsid can comprise a capsid created by capsid evolution or by rational capsid engineering from a naturally isolated AAV variants to capture desirable serotype features such as enhanced or specific tissue tropism and / or a host immune response escape. In an aspect, a disclosed evolved capsid can comprise AAVcc.47, AAVcc.81, or AAVcc.84. In an aspect, a disclosed AAV capsid can comprise the capsid of AAVrh39, AAVrh43, or AAVcy.7. In an aspect, a disclosed AAV capsid can comprise the capsid of a naturally isolated AAV variant. Variants include, but not limited to, AAV-DJ, AAV-DJ / 8, AAV- HAE1, AAV-HAE2, AAVM41, AAV-1829, AAV2 Y / F, AAV2 T / V, AAV2i8, AAV2.5, AAV2.7m8, AAV9.45, AAV9.61, AAV-B1, AAV-AS, AAV9.45A-String (e.g., AAV9.45-AS), AAV9.45Angiopep, AAV9.47-Angiopep, and AAV9.47-AS, AAV-PHP.B, AAV-PHP.eB, AAV-PHP.S, and AAV-F. In an aspect, a disclosed AAV capsid can comprise the capsid of AAV- Rh74 or a related variant (e.g., capsid variants like RHM4-1).2. Linkers

[0124] Disclosed herein is a linker. Disclosed herein is a linker that can be used to fuse a disclosed J peptide with a disclosed cargo. Disclosed herein is a linker that can be used to fuse a disclosed J peptide with a disclosed cargo. In an aspect, a disclosed linker can be fused to the N terminus of a disclosed peptide or a disclosed J peptide. In an aspect, a disclosed linker can be fused to the C terminus of a disclosed peptide or a disclosed J peptide. In an aspect, a disclosed linker can a flexible linker or a rigid linker. In an aspect, a disclosed linker can a Whitlow linker. In an aspect, a disclosed linker can comprise the sequence of SEQ ID NO: 185 or SEQ ID NO: 186. In an aspect, a disclosed linker can comprise the sequence of any one of SEQ ID NO: 187 - SEQ ID NO: 192.

[0125] In an aspect, a disclosed peptide can be linked to or fused with a disclosed cargo. In an aspect, a disclosed peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid can be linked to or fused with a disclosed cargo. In an aspect, a disclosed peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid can be linked to or fused with a disclosed cargo. In an aspect, a disclosed peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid can be linked to or fused with a disclosed cargo. In an aspect, a disclosed peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid can be linked to fused with a disclosed cargo.Table 2 - Listing of Exemplary Linkers

[0126] In an aspect, a disclosed cargo can be linked to or fused with the N-terminus of a disclosed peptide. In an aspect, a disclosed cargo can be linked to or fused with the C-terminus of a disclosed peptide. In an aspect, a disclosed cargo can be linked to or fused with the N-terminus of a disclosed J peptide. In an aspect, a disclosed cargo can be linked to or fused with the C-terminus of a disclosed J peptide.

[0127] In an aspect, a disclosed linker can comprise the sequence set forth in any one of SEQ ID NO: 193 - SEQ ID NO:213. In an aspect, a disclosed linker can comprise a sequence having at least 90% identity to the sequence set forth in any one of SEQ ID NO: 193 - SEQ ID NO:213.3. Nucleic Acid Molecules

[0128] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid.

[0129] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid.

[0130] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid.

[0131] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid.

[0132] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV)capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0133] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV), wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0134] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV), wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0135] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinantadeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11.

[0136] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:11. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:11. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11.

[0137] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0138] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0139] In an aspect, a disclosed encoded peptide can be referred to as a “J peptide”. In an aspect, a disclosed encoded peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid can be referred to as a “J peptide”. In an aspect, a disclosed encoded peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) can be referred to as a “J peptide”. In an aspect, a disclosed encoded peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid can be referred to as a “J peptide”. In an aspect, a disclosed encoded peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) can be referred to as a “J peptide”.

[0140] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associatedvirus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98.

[0141] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64.

[0142] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60.

[0143] Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encodinga peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78. Disclosed herein is a nucleic acid molecule comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the nucleic acid sequence comprises the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78.

[0144] In an aspect, a disclosed nucleic acid sequence can comprise the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. In an aspect, a disclosed nucleic acid sequence can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. In an aspect, a disclosed nucleic acid sequence can comprise the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. In an aspect, a disclosed nucleic acid sequence can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. In an aspect, a disclosed nucleic acid sequence can comprise the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. In an aspect, a disclosed nucleic acid sequence can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. In an aspect, a disclosed nucleic acid sequence can comprise the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78. In an aspect, a disclosed nucleic acid sequence can comprise a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78.

[0145] In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:50 can encode the sequence set forth in SEQ ID NO:01. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:51 can encode the sequence set forth in SEQ ID NO:02. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:52 can encode the sequence set forth in SEQ ID NO:03. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:53 can encode the sequence set forth in SEQ ID NO:04. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO: 54 can encode the sequence set forth in SEQ ID NO: 05. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:55can encode the sequence set forth in SEQ ID NO:06. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:56 can encode the sequence set forth in SEQ ID NO:07. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:57 can encode the sequence set forth in SEQ ID NO:08. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:58 can encode the sequence set forth in SEQ ID NO:09. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:59 can encode the sequence set forth in SEQ ID NO: 10. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:60 can encode the sequence set forth in SEQ ID NO: 11. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:61 can encode the sequence set forth in SEQ ID NO: 12. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:62 can encode the sequence set forth in SEQ ID NO: 13. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:63 can encode the sequence set forth in SEQ ID NO: 14. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:64 can encode the sequence set forth in SEQ ID NO: 15. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:65 can encode the sequence set forth in SEQ ID NO: 16. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:66 can encode the sequence set forth in SEQ ID NO: 17. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:67 can encode the sequence set forth in SEQ ID NO: 18. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:68 can encode the sequence set forth in SEQ ID NO: 19. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:69 can encode the sequence set forth in SEQ ID NO:20. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:70 can encode the sequence set forth in SEQ ID NO:21. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:71 can encode the sequence set forth in SEQ ID NO:22. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:72 can encode the sequence set forth in SEQ ID NO:23. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:73 can encode the sequence set forth in SEQ ID NO:24. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:74 can encode the sequence set forth in SEQ ID NO:25. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:75 can encode the sequence set forth in SEQ ID NO:26. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:76 can encode the sequence set forth in SEQ ID NO:27. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:77 can encode the sequence set forth in SEQ IDNO:28. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:78 can encode the sequence set forth in SEQ ID NO:29. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:79 can encode the sequence set forth in SEQ ID NO:30. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:80 can encode the sequence set forth in SEQ ID NO:31. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:81 can encode the sequence set forth in SEQ ID NO:32. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:82 can encode the sequence set forth in SEQ ID NO:33. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:83 can encode the sequence set forth in SEQ ID NO:34. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:84 can encode the sequence set forth in SEQ ID NO:35. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:85 can encode the sequence set forth in SEQ ID NO:36. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:86 can encode the sequence set forth in SEQ ID NO:37. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO: 87 can encode the sequence set forth in SEQ ID NO:38. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:88 can encode the sequence set forth in SEQ ID NO:39. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:89 can encode the sequence set forth in SEQ ID NO:40. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:90 can encode the sequence set forth in SEQ ID NON 1. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:91 can encode the sequence set forth in SEQ ID NO:42. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:92 can encode the sequence set forth in SEQ ID NO:43. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:93 can encode the sequence set forth in SEQ ID NO:44. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:94 can encode the sequence set forth in SEQ ID NO:45. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:95 can encode the sequence set forth in SEQ ID NO:46. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:96 can encode the sequence set forth in SEQ ID NO:47. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:97 can encode the sequence set forth in SEQ ID NO:48. In an aspect, a disclosed nucleotide sequence comprising the sequence set forth in SEQ ID NO:98 can encode the sequence set forth in SEQ ID NO:49.Table 3 - Listing of Exemplary J Peptides

[0146] Disclosed herein is a nucleic acid sequence encoding a peptide fused to a linker. Disclosed herein is a nucleic acid sequence encoding a J peptide fused to a linker. In an aspect, a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126. In an aspect, a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126. In an aspect, a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126 and can be linked to a disclosed cargo. In an aspect, a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, ormore than about 99% identity to the sequence set forth in any one of SEQ ID NO: 99 - SEQ ID NO: 126 and can be linked to a disclosed cargo.

[0147] In an aspect, a disclosed nucleic acid molecule can further comprise a nuclear localization signal (NLS). In an aspect, a disclosed NLS can comprise any NLS known to the art. As known to the art, nuclear localization signals (NLS) are generally short peptides that act as a signal fragment that mediates the transport of proteins from the cytoplasm into the nucleus.

[0148] In an aspect, a disclosed nucleic acid sequence can encode the protein or a portion thereof (such as, for example, Exon 1 or Exon 4, etc.) associated with the following genes: ABCA1, ABCA12, ABCA13, ABCA2, ABCA3, ABCA4, ABCA5, ABCC1, ABCC2, ABCC6, ABCC8, ABCC9, ACAN, ADAMTS13, ADCY10, ADGRV1, AGL, AGRN, AHDC1, ALK, ALMS1, ALPK3, ALS2, ANAPC1, ANK1, ANK2, ANK3, ANKRD11, ANKRD26, APC, APC2, APOB, ARFGEF2, ARHGAP31, ARHGEF10, ARHGEF18, ARID1A, ARID I B, ARID2, ASH1L, ASPM, ASXL1, ASXL2, ASXL3, ATM, ATP7A, ATP7B, ATR, ATRX, BAZ1A, BAZ2B, BCOR, BCORL1, BDP1, BLM, BPTF, BRCA1, BRCA2, BRD4, BRWD3, C2CD3, C3, C5, CACNA1A, CACNA1B, CACNA1C, CACNA1D, CACNA1E, CACNA1F, CACNA1G, CACNA1H, CACNA1S, CAD, CAMTAI, CARMIL2, CC2D2A, CCDC88A, CCDC88C, CCNB3, CDH23, CDK13, CDK5RAP2, CELSR1, CEMIP2, CENPE, CENPF, CENPJ, CEP152, CEP164, CEP250, CEP290, CFAP43, CFAP44, CFAP65, CFTR / ABCC7, CHD1, CHD2, CHD3, CHD4, CHD7, CHD8, CIC, CIT, CLIP1, CLTC, CNOT1, CNTNAP1, COL11A1, COL11A2, COL12A1, COL17A1, COL18A1, COL1A1, COL1A2, COL27A1, COL2A1, COL3A1, COL4A1, COL4A2, COL4A3, COL4A4, COL4A5, COL4A6, COL5A1, COL5A2, COL6A3, COL7A1, CPAMD8, CPLANE1, CPS1, CPSF1, CRB1, CREBBP, CUBN, CUL7, CUX1, DCC, DCHS1, DEPDC5, DICER1, DIP2B, DLC1, DMD, DMXL2, DNAH1, DNAH11, DNAH17, DNAH2, DNAH5, DNAH7, DNAH8, DNAH9, DNMBP, DNMT1, DOCK2, DOCK3, DOCK6, DOCK7, DOCK8, DSCAM, DSP, DST, DUOX2, DYNC1H1, DYNC2H1, DYSF, EIF2AK4, EP300, EPG5, ERCC6, ERCC6L2, EXPH5, EYS, F5, F8, FANCA, FANCD2, FANCM, FAT1, FAT4, FBN1, FBN2, FLG, FLG2, FLNA, FLNB, FLNC, FLT4, FMN2, FN1, FRAS1, FREM1, FREM2, FSIP2, FYCO1, GLI2, GLI3, GPR179, GREB1L, GRIN2A, GRIN2B, GRIN2D, HCFC1, HECW2, HERC1, HERC2, HFM1, HIVEP1, HIVEP2, HMCN1, HSPG2, HTT, HUWE1, HYDIN, IFT140, IFT172, IGF1R, IGF2R, IGSF1, INSR, INTS1, IQSEC2, ITGB4, ITPR1, ITPR2, JMJD1C, KALRN, KANK1, KAT6A, KAT6B, KDM3B, KDM5B, KDM5C, KDM6A, KDM6B, KDR, KIAA0586, KIAA1109, KIAA1549, KIDINS220, KIF14, KIF1A, KIF1B, KIF21A, KIF26B, KIF7, KMT2A, KMT2B, KMT2C, KMT2D, KMT2E, KNL1, LAMA1, LAMA2, LAMA3, LAMA4, LAMA5, LAMB1, LAMB2, LAMC3, LCT, LOXHD1, LPA, LRBA, LRP1, LRP2, LRP4, LRP5, LRP6, LRPPRC, LRRK1, LRRK2, LTBP2, LTBP4,LYST, MACF1, MADD, MAGI2, MAP1B, MAP3K1, MAPK8IP3, MAPKBP1, MAST1, MBD5, MCM3AP, MED12, MED12L, MED13, MED13L, MED23, MEGF8, MET, MLH3, MPDZ, MSH6, MTOR, MYH10, MYH11, MYH14, MYH2, MYH3, MYH6, MYH7, MYH7B, MYH8, MYH9, MYLK, MYO 15 A, MYO18B, MYO3A, MYO5A, MYO5B, MYO7A, MYO9A, NALCN, NBAS, NBEA, NBEAL2, NCAPD2, NCAPD3, NEB, NEXMIF, NEXMIF, NF1, NFASC, NHS, NIN, NIPBL, NLRP1, NOTCH1, NOTCH2, NOTCH3, NPHP4, NRXN1, NRXN3, NSD1, NSD2, NUP155, NUP188, NUP205, OBSCN, OBSL1, OTOF, OTOG, OTOGL, PARD3, PBRM1, PCDH15, PCLO, PCNT, PHIP, PI4KA, PIEZO1, PIEZO2, PIK3C2A, PIKFYVE, PKD1, PKD1L1, PKHD1, PLCE1, PLEC, PLEKHG2, PNPLA6, POGZ, POLA1, POLE, POLR1A, POLR2A, POLR3A, PRG4, PRKDC, PRPF8, PRR12, PRX, PTCHI, PTPN23, PTPRF, PTPRJ, PTPRQ, PXDN, QRICH2, RAB3GAP2, RAH, RALGAPA1, RANBP2, RB1CC1, RELN, RERE, REV3L, RIC1, RIMS1, RIMS2, RNF213, ROBO1, ROBO2, ROBO3, ROS1, RP1, RP1L1, RTTN, RUSC2, RYR1, RYR2, SACS, SAMD9, SAMD9L, SBF2, SCAPER, SCN10A, SCN11A, SCN1A, SCN2A, SCN3A, SCN4A, SCN5A, SCN8A, SCN9A, SETBP1, SETD1A, SETD1B, SETD2, SETD5, SETX, SHANK2, SHANK3, SHROOM4, SI, SIPA1L3, SLIT2, SLX4, SMARCA2, SMARCA4, SMCHD1, SNRNP200, SON, SPEF2, SPEG, SPG11, SPTA1, SPTAN1, SPTB, SPTBN2, SPTBN4, SRCAP, STRC, SVIL, SYNE1, SYNGAP1, SYNJ1, SZT2, TAF1, TANC2, TCF20, TCOF1, TDRD9, TECPR2, TECTA, TENM3, TENM4, TET3, TEX14, TEX15, TG, THOC2, TMEM94, TNC, TNIK, TNR, TNRC6B, TNXB, TOGARAMI, TONSL, TRIO, TRIOBP, TRIP11, TRIP12, TRPM1, TRPM6, TRPM7, TRRAP, TSC2, TTC37, TTN, TUBGCP6, UBR1, UNC80, USH2A, USP9X, VCAN, VPS13A, VPS13B, VPS13C, VPS13D, VWF, WDFY3, WDR19, WDR62, WDR81, WNK1, WRN, ZFHX2, ZFYVE26, ZNF142, ZNF292, ZNF335, ZNF407, ZNF462, ZNF469, or a portion thereof.

[0149] In an aspect, a disclosed nucleic acid sequence to be trans-spliced can be CpG depleted and codon-optimized for expression in a human cell. In an aspect, “CpG-free” can mean completely free of CpGs or partially free of CpGs. In an aspect, “CpG-free” can mean “CpG- depleted”. In an aspect, “CpG-depleted” can mean “CpG-free”. In an aspect, “CpG-depleted” can mean completely depleted of CpGs or partially depleted of CpGs. In an aspect, “CpG-free” can mean “CpG-optimized” for a desired and / or ideal expression level. CpG depletion and / or optimization is known to the skilled person in the art.4. Cargo

[0150] Disclosed herein is a cargo that is linked to or fused with a disclosed peptide or a disclosed J peptide. In an aspect, a disclosed cargo can be linked to or fused with the N-terminus of a disclosed peptide. In an aspect, a disclosed cargo can be linked to or fused with the C-terminusof a disclosed peptide. In an aspect, a disclosed cargo can be linked to or fused with the N- terminus of a disclosed J peptide. In an aspect, a disclosed cargo can be linked to or fused with the C-terminus of a disclosed J peptide.

[0151] In an aspect, a disclosed linker can a flexible linker or a rigid linker. In an aspect, a disclosed linker can a Whitlow linker. In an aspect, a disclosed linker can comprise the sequence of SEQ ID NO: 185 or SEQ ID NO: 186. In an aspect, a disclosed linker can comprise the sequence of any one of SEQ ID NO: 187 - SEQ ID NO: 192. In an aspect, a disclosed linker can be encoded by the sequence set forth in any one of SEQ ID NO: 193 - SEQ ID NO:213.

[0152] In an aspect, a disclosed J peptide can be linked to or fused with a disclosed cargo. In an aspect, a disclosed peptide can be linked to or fused with a disclosed cargo.

[0153] In an aspect, a disclosed cargo can be linked to or fused with a disclosed peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cargo can be linked to or fused with a disclosed peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cargo can be linked to or fused with a disclosed peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid. In an aspect, a disclosed cargo can be linked to fused with a disclosed peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid.

[0154] In an aspect, a disclosed cargo can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme. In an aspect, a disclosed cargo can comprise restoring one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, restoring one or more aspect of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation comprises restoring the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme. In an aspect of a disclosed cargo, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell types; (ii) normalizing aspects of the autophagy pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or preserving organelle functionality and / or structural integrity; (v) correcting enzyme dysregulation; (vi) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the multi -systemic manifestations of a genetic disease or disorder; (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of a genetic disease or disorder, or (viii) any combination thereof.

[0155] In an aspect, restoring one or more aspects of cellular homeostasis can comprise improving, enhancing, restoring, and / or preserving one or more aspects of cellular structural and / or functional integrity. In an aspect, restoring the activity and / or functionality of a missing, deficient, and / or mutant protein or enzyme can comprise a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10-20%, 20- 30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a control level or a reference level (e.g., determined, for example, using one or more subjects not having a missing, deficient, and / or mutant protein or enzyme). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity, and / or functionality is similar to that of a wild-type or control level.

[0156] In an aspect, a disclosed cargo can comprise a protein or a therapeutic protein. In an aspect, a disclosed cargo can comprise an enzyme or a therapeutic enzyme.

[0157] In an aspect, a disclosed cargo can comprise a therapeutic cargo. In an aspect, a disclosed therapeutic cargo can comprise a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject.

[0158] In an aspect, a disclosed therapeutic cargo can replace and / or supplement a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject.

[0159] In an aspect, a disclosed cargo can be a non-DNA cargo.

[0160] In an aspect, a disclosed cargo can comprise an RNA binder. In an aspect, a disclosed RNA binder can comprise a small molecule that binds to a target RNA. In an aspect, a disclosed RNA binder can comprise a macrolide, an alkaloid, an aminoglycoside, an anthracene, an oxazolidinone, a ribocil, an SMN2 ligand, a tetracycline, or a triptycene.

[0161] In an aspect, a disclosed small molecule that binds to a target RNA can be an anthracene, a kanamycin, a linezolid, a neomycin, a paromomycin, a pleuromutilin, a ribocil, a tedizolid, or a triptycene. In an aspect, a disclosed RNA binder can comprise a CPMV arginine rich motif (ARM), a STNV arginine rich motif (ARM), LambdaN, or a MS2.

[0162] In an aspect, a disclosed RNA binder can comprise roseoflavin. In an aspect, a disclosed cargo can comprise a molecular glue (MG). In an aspect, a disclosed MG can comprise cyclosporin, FK506, thalidomide, lenalidomide, or any combination thereof. In an aspect, a disclosed molecular glue can be a small molecules that possess intrinsic property to induce interaction between two proteins. In an aspect, a disclosed MG can promote the dimerization or colocalization between two unrelated proteins via formation of a ternary complex. In an aspect,these induced interactions can be between a target protein and a ubiquitin ligase, and can result in its proteasomal degradation or between two unrelated proteins and a perturbation of signaling cascades.

[0163] In an aspect, a disclosed cargo can comprise an epigenetic modulator for activation or repression. In an aspect, a disclosed epigenetic modulator can comprise a polypeptide having some type of activation activity or repression activity. In an aspect, a disclosed epigenetic modulator can comprise a DNA methyltransferase (DNMT) inhibitor, a histone methyltransferase, a demethylase, an acetyltransferase, a deacetylase, a bromodomain and extra-terminal domain (BET) inhibitor, any fusion thereof, or any combination thereof. In an aspect, a disclosed epigenetic modulator can comprise a polypeptide having transcription activation activity, transcription repression activity, transcription release factor activity, histone modification activity, nucleic acid association activity, methyltransferase activity, demethylase activity, acetyltransferase activity, deacetylase activity, or any combination thereof. In an aspect, a disclosed encoded polypeptide can comprise HP la, HP lb, MBD1, MBD2, Kriippel-Associated Box (KRAB), NIPP1, the Transcription Repression Domain (TRD) of Methyl-CpG Binding Protein 2 (MeCP2), DNMT3A, any fusion thereof, or any combination thereof. In an aspect, a disclosed fusion or disclosed fusion protein can comprise HPla-HPla, HPla-HPlb, HPla-MBDl, HPla-MBD2, HPla-KRAB, HPla-NIPPl, HPla-MeCP2, HPla-DNMT3A, HPlb-HPlb, HPlb- HPla, HPlb-MBDl, HPlb-MBD2, HPlb-KRAB, HPlb-NIPPl, HPlb-MeCP2, HPlb- DNMT3A, MBD1-MBD1, MBDl-HPla, MBDl-HPlb, MBD1-MBD2, MBD1-KRAB, MBD1- NIPP1, MBDl-MeCP2, MBD1-DNMT3A, MBD2-MBD2, MBD2-HPla, MBD2-HPlb, MBD2- MBD1, MBD2-KRAB, MBD2-NLPP1, MBD2-MeCP2, MBD2-DNMT3A, KRAB-KRAB, KRAB-HPla, KRAB-HPlb, KRAB-MBD1, KRAB-MBD2, KRAB-NIPP1, KRAB-MeCP2, KRAB-DNMT3A, NIPPI-NIPPI, NIPPl-HPla, NIPPl-HPlb, NIPP1-MBD1, NIPP1-MBD2, NIPP1-KRAB, NIPPl-MeCP2, NIPP1-DNMT3A, MeCP2-MeCP2, MeCP2-HPla, MeCP2- HPlb, MeCP2-MBDl, MeCP2-MBD2, MeCP2-KRAB, MeCP2-NIPPl, MeCP2-DNMT3A, DNMT3A-DNMT3A, DNMT3A-HPla, DNMT3A-HPlb, DNMT3A-MBD1, DNMT3A-MBD2, DNMT3A-KRAB, DNMT3A-NIPP1, or DNMT3 A-MeCP2.

[0164] In an aspect, a disclosed cargo can comprise a transcription factor or a binder thereof. In an aspect, exemplary transcription factors can include, but are not limited to, nuclear factor KB (NF-KB) (e.g., 50, p52, c-Rel, p65 (Rel-A), and Rel-B), activating protein 1 (AP-1), signal transducer and activator of transcription factor (STAT)-3, and members of the CCAAT / enhancer- binding protein (CZEBP) family of transcription factors (e.g., CZEBPP and CZEBP5).

[0165] As known in the art, transcription factors (TFs) recognize specific DNA sequences to control chromatin and transcription, forming a complex system that guides expression of thegenome. As reviewed by Lambert SA et al. (2018), there are over 1600 likely human TFs and binding motifs for two-thirds of them. TFs cannot be understood functionally without accompanying detailed knowledge of the DNA sequences they bind. TF DNA-binding specificities are frequently summarized as “motifs” - models representing the set of related, short sequences preferred by a given TF, which can be used to scan longer sequences (e.g., promoters) to identify potential binding sites. Determining a DNA-binding motif is often the first step toward detailed examination of the function of a TF because identification of potential binding sites provides a gateway to further analyses.

[0166] In an aspect, a disclosed cargo can comprise an inhibitor of a cellular degradation process. In an aspect, a disclosed inhibitor of a cellular degradation process can comprise a protease blocker or inhibitor and / or a proteosome blocker or inhibitor. In an aspect, a disclosed protease and / or proteasome blocker or inhibitor can comprise Pepstatin A (can inhibit aspartic acid proteases), Leupeptin (can inhibit serine and cysteine proteases), EDTA (can inhibit metalloproteases), PMSF (can inhibit serine proteases), Bortezomib (e.g., a proteasome inhibitor that is a slowly reversible inhibitor), Carfilzomib (e.g., a proteasome inhibitor that is an irreversible inhibitor), Delanzomib, Ixazomib (e.g., a proteasome inhibitor that is a reversible inhibitor), or any combination thereof. In an aspect, a disclosed inhibitor of a cellular degradation process can comprise an autophagy inhibitor. In an aspect, a disclosed autophagy inhibitor can comprise E64d (e.g., a membrane- permeable inhibitor of cathepsins B, H, and L), Pepstatin A (can inhibit cathepsins D and E), Baf Al (can inhibit vacuolar H+ ATPase), Monensin (can mediate the exchange of protons for potassium or sodium), 3 -MA (can prevent the starvation-induced colocalization of mitochondrial and lysosomal markers), or any combination thereof.

[0167] In an aspect, a disclosed cargo can comprise a DNA binder. In an aspect, a disclosed DNA binder can be a small molecule (can bind to DNA through intercalation or minor groove binding), a DNA-binding protein (can bind to DNA through domains such as helix-turn-helix, zinc finger, and homeodomains), a bisbenzamidine (can bind to the minor groove of DNA), a photosensitive metal complex (can bind to DNA duplexes), or any combination thereof.

[0168] In an aspect, a disclosed DNA binder can alter the stability and functionality of DNA, can be used to program reaction pathways of dynamic DNA reactions, can be used to trigger site- selective, DNA-promoted biochemical and physicochemical processes, or any combination thereof. In an aspect, a disclosed DNA binder can be used to accelerate the discovery and characterization of bioactive small molecules, to develop high-throughput screening assays for discovering DNA binders, or any combination thereof. In an aspect, a disclosed DNA binder can bind to DNA, thereby altering the structure and / or function of DNA.

[0169] In an aspect, a disclosed DNA binding protein can comprise a polypeptide motif that enables the binding to the major groove of DNA. In an aspect, a disclosed motif can comprise helix-turn-helix (HTH), helix-loop-helix (HLH), zinc fingers, and leucine zippers. HTH and HLH are similar in that they both contain two a-helices separated by a turn (short) and a loop (long) region. Both usually bind as dimers to inverted repeats in the DNA sequence.

[0170] Leucine zippers are a-helices that contain a leucine residue every seventh amino acid. This motif is found in many eukaryotic transcription factors. Zinc fingers consist of 25-30 amino acids surrounding a single zinc atom, which is coordinated by two cysteines, which are very close to short a-helices. In an aspect, a disclosed helix can contact the DNA.

[0171] In an aspect, a disclosed DNA binder can comprise a bisbenzamidine or a pyrazine. In an aspect, a disclosed DNA binder can comprise a HuH or a dead HuH, an arginine rich motif (ARM) (e.g., a MSV ARM), an HIV enhancer-binding peptide R42, a yeast transcriptional activator GCN4, a major break region (MBR) (e.g., MBR-CC).

[0172] In an aspect, a disclosed cargo can comprise a CRISPR / Cas protein. In an aspect, a disclosed CRISPR / Cas protein can comprise a CRISPR-based endonuclease. In an aspect, a disclosed CRISPR-based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system. In an aspect, a disclosed therapeutic agent can comprise a CRISPR-based endonuclease (e.g., Cas9 or Casl3). In an aspect, a disclosed CRISPR-based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system. In aspect, Cas9 (e.g., SpCas9 and SaCas9) are well-known to the art. In aspect, Casl3 comprises at least four known subtypes, including Cast 3a (formerly C2c2), Cast 3b, Cast 3 c, and Cast 3d. All known Cast 3 family members contain two HEPN domains, which confer RNase activity. In an aspect, Cast 3 can be considered an outlier in the CRISPR world because it targets RNA, not DNA. Once it is activated by a ssRNA sequence bearing complementarity to its crRNA spacer, it unleashes a nonspecific RNase activity and destroys all nearby RNA regardless of their sequence. As disclosed herein, this property can be harnessed in vitro for precision diagnostics. Generally, Cast 3 can be found in Leptotrichia buccalis, Leptotrichia shahii, Ruminococcus flavefaciens, Bergeyella zoohelcum, Prevotella buccae, and Listeria seeligeri and can have a size of about 900 to about 1300 amino acids. In an aspect, the guide spacer length can be about 22 to about 30 nucleotides while the total guide length can be about 52 to about 66 nucleotides. In an aspect, a PAM can be 3-H for LshCasl3a, 5-D and 3-NAN or NNA for BzCasl3b, and none for RfCasl3d. In an aspect, a disclosed Casl3 can cut ssRNA.

[0173] As known to the skilled person in the cart, a Casl3d ortholog can be from a prokaryotic genome or metagenome, gut metagenome, an activated sludge metagenome, an anaerobic digester metagenome, a chicken gut metagenome, a human gut metagenome, a pig gut metagenome, abovine gut metagenome, a sheep gut metagenome, a goat gut metagenome, a capybara gut metagenome, a primate gut metagenome, a termite gut metagenome, a fecal metagenome, a genome from the Order Clostridiales, or the Family Ruminococcaceae. In an aspect, a disclosed Cast 3d ortholog can include an Cast 3d ortholog from Ruminococcus albus, Eubacterium siraeum, a Ruminococcus flavefaciens strain XPD3002, Ruminococcus flavefaciens FD-1, uncultured Eubacterium sp TS28-c4095, uncultured Ruminococcus sp., Ruminococcus bicirculans, or Ruminococcus sp CAG57.

[0174] In an aspect, a disclosed CRISPR / Cas protein can comprise CRISPR-Cas (e.g., a small RNA-guided enzyme found uniquely in bacteriophages that can achieve programmable DNA cutting as well as genome editing).

[0175] In an aspect, a disclosed CRISPR / Cas protein can comprise a guide RNA (gRNA). In an aspect, a disclosed gRNA can comprise a specific RNA sequence that recognizes the target DNA region of interest and can direct a disclosed Cas nuclease there for editing. In an aspect, a disclosed gRNA can comprise 2 parts: (i) crispr RNA (crRNA), which is a 17-20 nucleotide sequence complementary to the targeted DNA or targeted locus, and (ii) a tracr RNA, which serves as a binding scaffold for a Cas nuclease. In an aspect, a targeted DNA or targeted locus can be a gene having one or more mutations or defects that contribute to one or more genetic diseases or disorders. In an aspect, a targeted DNA or targeted locus can be subjected to a Cas9 induced double-stranded break.

[0176] In an aspect, a disclosed tracrRNA sequence can comprise nucleotides that hybridize to a minimum CRISPR repeat sequence in a cell. A minimum tracrRNA sequence and a minimum CRISPR repeat sequence may form a duplex, i.e., a base-paired double-stranded structure. Together, the minimum tracrRNA sequence and the minimum CRISPR repeat can bind to an RNA-guided endonuclease. At least a part of the minimum tracrRNA sequence can hybridize to the minimum CRISPR repeat sequence. In an aspect, a disclosed minimum tracrRNA sequence can be at least about 30%, about 40%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or 100% complementary to the minimum CRISPR repeat sequence. For example, in an aspect, a disclosed minimum tracrRNA sequence can have a length from about 7 nucleotides to about 100 nucleotides. In an aspect, a disclosed minimum tracrRNA sequence can be from about 7 nucleotides (nt) to about 50 nt, from about 7 nt to about 40 nt, from about 7 nt to about 30 nt, from about 7 nt to about 25 nt, from about 7 nt to about 20 nt, from about 7 nt to about 15 nt, from about 8 nt to about 40 nt, from about 8 nt to about 30 nt, from about 8 nt to about 25 nt, from about 8 nt to about 20 nt, from about 8 nt to about 15 nt, from about 15 nt to about 100 nt, from about 15 nt to about 80 nt, from about 15 nt to about 50 nt, from about 15 nt to about 40 nt, from about 15 nt to about 30 nt or from about 15 nt to about 25 nt long.In an aspect, a disclosed minimum tracrRNA sequence can be at least about 60% identical to a reference minimum tracrRNA (e.g. wild type, tracrRNA from S. pyogenes) sequence over a stretch of at least 6, 7, or 8 contiguous nucleotides. In an aspect, for example, the minimum tracrRNA sequence can be at least about 65% identical, about 70% identical, about 75% identical, about 80% identical, about 85% identical, about 90% identical, about 95% identical, about 98% identical, about 99% identical or 100% identical to a reference minimum tracrRNA sequence over a stretch of at least 6, 7, or 8 contiguous nucleotides.

[0177] In an aspect, a disclosed gRNA can target a PAM sequence at or near the targeted DNA or targeted locus. In an aspect, a spacer sequence can be designed to hybridize to a target polynucleotide that is located 5’ of a PAM of a disclosed Cas9 endonuclease. In an aspect, a disclosed spacer can match the targeted sequence or can have mismatches. In an aspect, S. pyogenes Cas9 recognizes a PAM that comprises the sequence 5’-NRG-3’, where R comprises either A or G, where N is any nucleotide and N is immediately 3’ of the target nucleic acid sequence targeted by the spacer sequence. In an aspect, S. aureus Cas9 recognizes a PAM that comprises the sequence 5'-NNGRRT-3' (where R represents A or G) an NN is immediately 3’ of the target nucleic acid sequence targeted by the spacer sequence. PAMs are known to the skilled person in the art. In an aspect, a disclosed gRNA can target a PAM sequence at or near any mutation or defect in a gene having one or more mutations or defects that contribute to one or more genetic diseases or disorders.

[0178] In an aspect, a disclosed cargo can comprise an oligonucleotide agent or an oligonucleotide therapeutic agent. In an aspect, a disclosed oligonucleotide agent or an oligonucleotide therapeutic agent can be a single-stranded or double-stranded DNA, iRNA, shRNA, siRNA, mRNA, non-coding RNA (ncRNA), an antisense molecule, miRNA, a morpholino, a peptidenucleic acid (PNA), or an analog or conjugate thereof. In an aspect, a disclosed cargo can be an ASO or an RNAi.

[0179] In an aspect, a disclosed cargo can comprise ribosomal RNA (rRNA), transfer RNA (tRNA), heterogeneous nuclear RNA (hnRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), Piwi-interacting RNA (piRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), singe guide RNA (sgRNA), non-coding RNA (ncRNA), long non-coding RNA (IncRNA), 7SL, Xist, short enhancer RNA (eRNA), circular RNA, intergenic RNA, or any combination thereof. In an aspect, a disclosed RNA can comprise IncRNA, siRNA, shRNA, sgRNA, circular RNA, snoRNA, miRNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise a functional non-coding RNA element.

[0180] In an aspect, a disclosed cargo can be an RNA therapeutic (such as, for example, an oligonucleotide that targets RNA). In an aspect, a therapeutic RNA can comprise one or moreexpression sequences. As known to the art, expression sequences can comprise an RNAi, shRNA, mRNA, non-coding RNA (ncRNA), an antisense such as an antisense RNA, miRNA, morpholino oligonucleotide, peptide-nucleic acid (PNA) or ssDNA (with natural, and modified nucleotides, including but not limited to, LNA, BNA, 2’-0-Me-RNA, 2’-ME0-RNA, 2’-F-RNA), or analog or conjugate thereof. In an aspect, a disclosed therapeutic RNA can comprise one or more long non-coding RNA (IncRNA), such as, for example, a long intergenic non-coding RNA (lincRNA), pre-transcript, pre-miRNA, pre-mRNA, competing endogenous RNA (ceRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), pseudo-gene, rRNA, or tRNA. In an aspect, ncRNA can be pi wi -interacting RNA (piRNA), primary miRNA (pri-miRNA), or premature miRNA (pre-miRNA). In an aspect, a disclosed therapeutic RNA or an RNA therapeutic can comprise antisense oligonucleotides (ASOs) that inhibit mRNA translation, oligonucleotides that function via RNA interference (RNAi) pathway, RNA molecules that behave like enzymes (ribozymes), RNA oligonucleotides that bind to proteins and other cellular molecules, and ASOs that bind to mRNA and form a structure that is recognized by RNase H resulting in cleavage of the mRNA target. In an aspect, RNA therapeutics can comprise RNAi and ASOs that inhibit mRNA translation. Generally speaking, as known to the art, RNAi operates sequence specifically and post-transcriptionally by activating ribonucleases which, along with other enzymes and complexes, coordinately degrade the RNA after the original RNA target has been cut into smaller pieces while antisense oligonucleotides bind to their target nucleic acid via Watson-Crick base pairing, and inhibit or alter gene expression via steric hindrance, splicing alterations, initiation of target degradation, or other events.

[0181] In an aspect, a disclosed cargo can be an antibody or an antibody fragment. In an aspect, a disclosed antibody or antibody fragment can comprise a Fab, a Fab’, a F(ab’)2, a Fv, a single chain (scFv), a scFV mutant, a fusion protein comprising a disclosed antibody portion, and any other modified immunoglobulin molecule comprising an antigen recognition site. In an aspect, a disclosed antibody cargo can comprise abagovomab, abciximab, abituzumab, abrilumab, actoxumab, adalimumab, adecatumumab, aducanumab, afelimomab, afutuzumab, alacizumab pegol, alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab mafenatox, anetumab ravtansine, anifrolumab, anrukinzumab, apolizumab, arcitumomab, ascrinvacumab, aselizumab, atezolizumab, atinumab, atlizumab (tocilizumab), atorolimumab, bapineuzumab, basiliximab, bavituximab, bectumomab, begelomab, belimumab, benralizumab, bertilimumab, besilesomab, bevacizumab, bezlotoxumab, biciromab, bimagrumab, bimekizumab, bivatuzumab mertansine, blinatumomab, blosozumab, bococizumab, brentuxim abvedotin, briakinumab, brodalumab, brolucizumab, brontictuzumab, canakinumab, cantuzumab mertansine, cantuzumab ravtansine, caplacizumab, capromab pendetide, carlumab, catumaxomab, cbr96-doxorubicinimmunoconjugate, cedelizumab, certolizumab pegol, cetuximab, citatuzumab bogatox, cixutumumab, clazakizumab, clenoliximab, clivatuzumab tetraxetan, codrituzumab, coltuximab ravtansine, conatumumab, concizumab, crenezumab, dacetuzumab, daclizumab, dalotuzumab, dapirolizumab pegol, daratumumab, dectrekumab, demcizumab, denintuzumab mafodotin, denosumab, derlotuximab biotin, detumomab, dinutuximab, diridavumab, dorlimomab aritox, drozitumab, duligotumab, dupilumab, durvalumab, dusigitumab, ecromeximab, eculizumab, edobacomab, edrecolomab, efalizumab, efungumab, eldelumab, elgemtumab, elotuzumab, elsilimomab, emactuzumab, emibetuzumab, enavatuzumab, enfortumab vedotin, enlimomab pegol, enoblituzumab, enokizumab, enoticumab, ensituximab, epitumomab cituxetan, epratuzumab, erlizumab, ertumaxomab, etanercept, etaracizumab, etrolizumab, evinacumab, evolocumab, exbivirumab, fanolesomab, faralimomab, farletuzumab, fasinumab, felvizumab, fezakinumab, ficlatuzumab, figitumumab, firivumab, flanvotumab, fletikumab, fontolizumab, foralumab, foravirumab, fresolimumab, fulranumab, futuximab, galiximab, ganitumab, gantenerumab, gavilimomab, gemtuzumab ozogamicin, gevokizumab, girentuximab, glembatumumab vedotin, golimumab, gomiliximab, guselkumab, ibalizumab, ibritumomab tiuxetan, icrucumab, idarucizumab, igovomab, imalumab, imciromab, imgatuzumab, inclacumab, indatuximab ravtansine, indusatumab vedotin, infliximab, inolimomab, inotuzumab ozogamicin, intetumumab, ipilimumab, iratumumab, isatuximab, itolizumab, ixekizumab, keliximab, labetuzumab, lambrolizumab, lampalizumab, lebrikizumab, lemalesomab, lenzilumab, lerdelimumab, lexatumumab, libivirumab, lifastuzumab vedotin, ligelizumab, lilotomab satetraxetan, lintuzumab, lirilumab, lodelcizumab, lokivetmab, lorvotuzumab mertansine, lucatumumab, lulizumab pegol, lumiliximab, lumretuzumab, mapatumumab, margetuximab, maslimomab, matuzumab, mavrilimumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mirvetuximab soravtansine, mitumomab, mogamulizumab, morolimumab, motavizumab, moxetumomab pasudotox, muromonab-cd3, nacolomab tafenatox, namilumab, naptumomab estafenatox, narnatumab, natalizumab, nebacumab, necitumumab, nemolizumab, nerelimomab, nesvacumab, nimotuzumab, nivolumab, nofetumomab merpentan, obiltoxaximab, obinutuzumab, ocaratuzumab, ocrelizumab, odulimomab, ofatumumab, olaratumab, olokizumab, omalizumab, onartuzumab, ontuxizumab, opicinumab, oportuzumab monatox, oregovomab, orticumab, otelixizumab, otlertuzumab, oxelumab, ozanezumab, ozoralizumab, pagibaximab, palivizumab, panitumumab, pankomab, panobacumab, parsatuzumab, pascolizumab, pasotuxizumab, pateclizumab, patritumab, pembrolizumab, pemtumomab, perakizumab, pertuzumab, pexelizumab, pidilizumab, pinatuzumab vedotin, pintumomab, placulumab, polatuzumab vedotin, ponezumab, priliximab, pritoxaximab, pritumumab, quilizumab, racotumomab, radretumab, rafivirumab, ralpancizumab, ramucirumab, ranibizumab,raxibacumab, refanezumab, regavirumab, reslizumab, rilotumumab, rinucumab, rituximab, robatumumab, roledumab, romosozumab, rontalizumab, rovelizumab, ruplizumab, sacituzumab govitecan, samalizumab, sarilumab, satumomab pendetide, secukinumab, seribantumab, setoxaximab, sevirumab, sibrotuzumab, sifalimumab, siltuximab, simtuzumab, siplizumab, sirukumab, sofituzumab vedotin, solanezumab, solitomab, sonepcizumab, sontuzumab, stamulumab, sulesomab, suvizumab, tabalumab, tacatuzumab tetraxetan, tadocizumab, talizumab, tanezumab, taplitumomab paptox, tarextumab, tefibazumab, telimomab aritox, tenatumomab, teneliximab, teplizumab, teprotumumab, tesidolumab, tetulomab, ticilimumab, tigatuzumab, tildrakizumab, tocilizumab, toralizumab, tosatoxumab, tositumomab, tovetumab, tralokinumab, trastuzumab, tregalizumab, tremelimumab, trevogrumab, tucotuzumab celmoleukin, tuvirumab, ublituximab, ulocuplumab, urelumab, urtoxazumab, ustekinumab, vandortuzumab vedotin, vantictumab, vanucizumab, vapaliximab, varlilumab, vatelizumab, vedolizumab, veltuzumab, vepalimomab, vesencumab, visilizumab, volociximab, vorsetuzumab mafodotin, votumumab, zalutumumab, zanolimumab, zatuximab, ziralimumab, zolimomab aritox, or any combination thereof.5. Plasmids

[0182] Disclosed herein is a plasmid comprising one or more disclosed nucleic acid sequences. Disclosed here are plasmids used in methods of making a disclosed composition such as, for example, a disclosed J peptide, a disclosed cargo, a disclosed nucleic acid molecule, a disclosed vector, a disclosed AAV particle, a disclosed AAV-like particle, or a disclosed pharmaceutical formulation. Plasmids and using plasmids are known to the art. Disclosed herein is a plasmid comprising one or more disclosed nucleic acid molecules. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a disclosed peptide. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a disclosed J peptide.

[0183] Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO: 15. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO:11. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO:29. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO:49. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO:15. Disclosed herein is a plasmid comprising a nucleic acid sequenceencoding a J peptide comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO: 11. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO:29. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01-SEQ ID NO:49. Disclosed herein is a plasmid comprising a nucleic acid sequence comprising the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. Disclosed herein is a plasmid comprising a nucleic acid sequence comprising a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98.

[0184] Disclosed herein is a plasmid comprising a nucleic sequence for a disclosed peptide or a disclosed J peptide linked to or fused with a nucleic acid sequence for a disclosed cargo.

[0185] In an aspect, a disclosed cargo can comprise any disclosed cargo. In an aspect, a disclosed cargo can comprise a protein or a therapeutic protein. In an aspect, a disclosed cargo can comprise an enzyme or a therapeutic enzyme. In an aspect, a disclosed cargo can comprise a therapeutic cargo. In an aspect, a disclosed therapeutic cargo can comprise a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject. In an aspect, a disclosed cargo can be a non-DNA cargo. In an aspect, a disclosed cargo can comprise an RNA binder. In an aspect, a disclosed cargo can comprise an epigenetic modulator for activation or repression. In an aspect, a disclosed cargo can comprise a transcription factor or a binder thereof. In an aspect, a disclosed cargo can comprise an inhibitor of a cellular degradation process. In an aspect, a disclosed cargo can comprise a DNA binder. In an aspect, a disclosed cargo can comprise a CRISPR / Cas protein. In an aspect, a disclosed cargo can comprise an oligonucleotide agent or an oligonucleotide therapeutic agent.

[0186] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats(ITRs), wherein the disclosed encoded J peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NOT E In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0187] Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a peptide fused to a linker. Disclosed herein is a plasmid comprising a nucleic acid sequence encoding a J peptide fused to a linker. Disclosed herein is a plasmid comprising a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126. Disclosed herein is a plasmid comprising a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126. Disclosed herein is a plasmid comprising a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126 and can be linked to a disclosed cargo. Disclosed herein is a plasmid comprising a disclosed nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise asequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126 and can be linked to a disclosed cargo. Disclosed herein is a plasmid comprising a disclosed nucleic acid sequence having the sequence set forth in any one of SEQ ID NO: 193 - SEQ ID NO:213.

[0188] Triple transfection and methods to effect triple transfection are known to the art. Kits and components for triple transfection are commercially available. As known to the art, for example, AAV requires a helper plasmid containing genes from adenovirus that mediate AAV replication (i.e., E4, E2a and VA). In an aspect of a disclosed method, triple transfection can comprise using a pHelper plasmid, a plasmid comprising Rep / Cap, and a transfer plasmid comprising a disclosed J peptide or disclosed peptide fused to a disclosed cargo. In an aspect, generating AAV can occur in suspension cells. In an aspect of a disclosed method, suspension cells can be incubated. In an aspect, suspension cells (e.g., HEK cells, HEKG2 cells, etc.) and methods of incubating suspension cells. In an aspect, a disclosed method can comprise incubating suspension cells.

[0189] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a “J peptide”. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno- associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid.

[0190] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15.

[0191] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV), wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV), wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0192] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno- associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno- associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11.

[0193] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno- associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno- associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

[0194] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78.

[0195] In an aspect of a disclosed cis plasmid or a disclosed pCis, a disclosed cassette can comprise one or more regulatory elements. In an aspect, one or more disclosed regulatory elements can comprise an internal ribosomal entry site (IRES), a promoter, an enhancer, a promoter / enhancer, one or more inverted terminal repeats (ITRs), a poly adenylation sequence, a transcription termination signal, any combination thereof, or a combination of all disclosed regulatory elements.

[0196] In an aspect, a disclosed promoter can be operably linked to (i) a nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide, or (ii) a nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide linked to a disclosed cargo.

[0197] In an aspect, a disclosed promoter and / or enhancer can be positioned 5’ (upstream) or 3’ (downstream) of a disclosed nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide under its control. As is known in the art, the distance between the placement of a promoter and / or enhancer from a disclosed nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide variation can be accommodated without loss of function.

[0198] In an aspect, a disclosed plasmid can be used in a disclosed triple transfection method or a disclosed modified transfection method. For example, a disclosed plasmid can comprise a helper plasmid (pHelper), a trans-complementing plasmid (pTrans), or a cis plasmid (pCis).6. AAV Particles and AAV-Like Particles

[0199] Disclosed herein is an AAV particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is an AAV-like particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a plurality of AAV particles comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a plurality AAV-like particles comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is an AAV particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is an AAV-like particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a plurality of AAV particles comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a plurality AAV-like particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is an AAV particle comprising a disclosed cargo or disclosed cargos. Disclosed herein is an AAV-like particle comprising a disclosed cargo or disclosed cargos. Disclosed herein is a plurality of AAV particles comprising a disclosed cargo or disclosed cargos. Disclosed herein is a plurality AAV-like particles comprising a disclosed cargo or disclosed cargos.

[0200] In an aspect, a disclosed AAV-like particle can be non-infectious and unable to replicate due to the absence of a viral genome. In an aspect, a plurality of disclosed AAV-like particles can be non-infectious and unable to replicate due to the absence of a viral genome.

[0201] In an aspect, a disclosed AAV particle and / or a disclosed AAV-like particle comprising a disclosed cargo can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme. In an aspect, a disclosed AAV particle and / or a disclosed AAV- like particle comprising a disclosed cargo can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, restoring one or more aspect of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation comprises restoring the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell types; (ii) normalizing aspects of the autophagy pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or preserving organelle functionality and / or structural integrity; (v) correcting enzyme dysregulation; (vi) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate ofprogression of the multi -systemic manifestations of a genetic disease or disorder; (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of a genetic disease or disorder, or (viii) any combination thereof.

[0202] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, restoring the activity and / or functionality of a missing, deficient, and / or mutant protein or enzyme can comprise a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a control level or a reference level (e.g., determined, for example, using one or more subjects not having a missing, deficient, and / or mutant protein or enzyme). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity, and / or functionality is similar to that of a wild-type or control level.

[0203] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a protein or a therapeutic protein. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an enzyme or a therapeutic enzyme. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a therapeutic cargo. In an aspect, a disclosed therapeutic cargo can comprise a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed therapeutic cargo can replace and / or supplement a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject.

[0204] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a non-DNA target. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an RNA binder. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed RNA binder can comprise a small molecule that binds to a target RNA. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed RNA binder can comprise a macrolide, an alkaloid, an aminoglycoside, an anthracene, an oxazolidinone, a ribocil, an SMN2 ligand, a tetracycline, or a tripty cene.

[0205] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed small molecule that binds to a target RNA is an anthracene, a kanamycin, a linezolid, a neomycin, a paromomycin, a pleuromutilin, a ribocil, a tedizolid, or a triptycene.

[0206] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed RNA binder can comprise a CPMV arginine rich motif (ARM), a STNV arginine rich motif (ARM), LambdaN, or a MS2. In an aspect, a disclosed RNA binder can comprise roseoflavin.

[0207] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a molecular glue. In an aspect, a disclosed MG can comprise cyclosporin, FK506, thalidomide, lenalidomide, or any combination thereof.

[0208] In an aspect, a disclosed molecular glue (MG) can be a small molecule that possess intrinsic property to induce interaction between two proteins. In an aspect, a disclosed MG can promote the dimerization or colocalization between two unrelated proteins via formation of a ternary complex. In an aspect, these induced interactions can be between a target protein and a ubiquitin ligase, and can result in its proteasomal degradation or between two unrelated proteins and a perturbation of signaling cascades.

[0209] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an epigenetic modulator for activation or repression. In an aspect, a disclosed epigenetic modulator can comprise a polypeptide having some type of activation activity or repression activity. In an aspect, a disclosed epigenetic modulator can comprise a DNA methyltransferase (DNMT) inhibitor, a histone methyltransferase, a demethylase, an acetyltransferase, a deacetylase, a bromodomain and extra-terminal domain (BET) inhibitor, any fusion thereof, or any combination thereof.

[0210] In an aspect, a disclosed epigenetic modulator can comprise a polypeptide having transcription activation activity, transcription repression activity, transcription release factor activity, histone modification activity, nucleic acid association activity, methyltransferase activity, demethylase activity, acetyltransferase activity, deacetylase activity, or any combination thereof.

[0211] In an aspect, a disclosed encoded polypeptide can comprise HPla, HPlb, MBD1, MBD2, Kriippel -Associated Box (KRAB), NIPP1, the Transcription Repression Domain (TRD) of Methyl-CpG Binding Protein 2 (MeCP2), DNMT3A, any fusion thereof, or any combination thereof. In an aspect, a disclosed fusion or disclosed fusion protein can comprise HPla-HPla, HPla-HPlb, HPla-MBDl, HPla-MBD2, HPla-KRAB, HPla-NIPPl, HPla-MeCP2, HPla- DNMT3A, HPlb-HPlb, HPlb-HPla, HPlb-MBDl, HPlb-MBD2, HPlb-KRAB, HPlb-NIPPl, HPlb-MeCP2, HPlb-DNMT3A, MBD1-MBD1, MBDl-HPla, MBDl-HPlb, MBD1-MBD2, MBD1-KRAB, MBD1-NIPP1, MBDl-MeCP2, MBD1-DNMT3A, MBD2-MBD2, MBD2-HPla, MBD2-HPlb, MBD2-MBD1, MBD2-KRAB, MBD2-NIPP1, MBD2-MeCP2, MBD2- DNMT3A, KRAB-KRAB, KRAB-HPla, KRAB-HPlb, KRAB-MBD1, KRAB-MBD2, KRAB- NIPP1, KRAB-MeCP2, KRAB-DNMT3A, NIPPI-NIPPI, NIPPl-HPla, NIPPl-HPlb, NIPP1- MBD1, NIPP1-MBD2, NIPP1-KRAB, NIPPl-MeCP2, NIPP1-DNMT3A, MeCP2-MeCP2,MeCP2-HPla, MeCP2-HPlb, MeCP2-MBDl, MeCP2-MBD2, MeCP2-KRAB, MeCP2-NIPPl, MeCP2-DNMT3A, DNMT3A-DNMT3A, DNMT3A-HPla, DNMT3A-HPlb, DNMT3A- MBD1, DNMT3A-MBD2, DNMT3A-KRAB, DNMT3A-NIPP1, or DNMT3 A-MeCP2.

[0212] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a transcription factor or a binder thereof. In an aspect, exemplary transcription factors can include, but are not limited to, nuclear factor KB (NF-KB) (e.g., 50, p52, c-Rel, p65 (Rel-A), and Rel-B), activating protein 1 (AP-1), signal transducer and activator of transcription factor (STAT)-3, and members of the CCAAT / enhancer-binding protein (CZEBP) family of transcription factors (e.g., CZEBPP and CZEBP5).

[0213] As known in the art, transcription factors (TFs) recognize specific DNA sequences to control chromatin and transcription, forming a complex system that guides expression of the genome.

[0214] As reviewed by Lambert SA et al. (2018), there are over 1600 likely human TFs and binding motifs for two-thirds of them. TFs cannot be understood functionally without accompanying detailed knowledge of the DNA sequences they bind. TF DNA-binding specificities are frequently summarized as “motifs” - models representing the set of related, short sequences preferred by a given TF, which can be used to scan longer sequences (e.g., promoters) to identify potential binding sites. Determining a DNA-binding motif is often the first step toward detailed examination of the function of a TF because identification of potential binding sites provides a gateway to further analyses.

[0215] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an inhibitor of a cellular degradation process.

[0216] In an aspect, a disclosed inhibitor of a cellular degradation process can comprise a protease blocker or inhibitor and / or a proteosome blocker or inhibitor. In an aspect, a disclosed protease and / or proteasome blocker or inhibitor can comprise Pepstatin A (can Inhibit aspartic acid proteases), Leupeptin (can inhibit serine and cysteine proteases), EDTA (can inhibit metalloproteases), PMSF (can inhibit serine proteases), Bortezomib (e.g., a proteasome inhibitor that is a slowly reversible inhibitor), Carfilzomib (e.g., a proteasome inhibitor that is an irreversible inhibitor), Delanzomib, Ixazomib (e.g., a proteasome inhibitor that is a reversible inhibitor), or any combination thereof. In an aspect, a disclosed inhibitor of a cellular degradation process can comprise an autophagy inhibitor. In an aspect, a disclosed autophagy inhibitor can comprise E64d (e.g., a membrane-permeable inhibitor of cathepsins B, H, and L), Pepstatin A (can inhibit cathepsins D and E), Baf Al (can inhibit vacuolar H+ ATPase), Monensin (can mediate the exchange of protons for potassium or sodium), 3 -MA (can prevent the starvation- induced colocalization of mitochondrial and lysosomal markers), or any combination thereof.

[0217] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a DNA binder. In an aspect, a disclosed DNA binder can be a small molecule (can bind to DNA through intercalation or minor groove binding), a DNA-binding protein (can bind to DNA through domains such as helix-turn-helix, zinc finger, and homeodomains), a bisbenzamidine (can bind to the minor groove of DNA), a photosensitive metal complex (can bind to DNA duplexes), or any combination thereof.

[0218] In an aspect, a disclosed DNA binder can alter the stability and functionality of DNA, can be used to program reaction pathways of dynamic DNA reactions, can be used to trigger site- selective, DNA-promoted biochemical and physicochemical processes, or any combination thereof. In an aspect, a disclosed DNA binder can be used to accelerate the discovery and characterization of bioactive small molecules, to develop high-throughput screening assays for discovering DNA binders, or any combination thereof.

[0219] In an aspect, a disclosed DNA binder can bind to DNA, thereby altering the structure and / or function of DNA. In an aspect, a disclosed DNA binding protein can comprise a polypeptide motif that enables the binding to the major groove of DNA. In an aspect, a disclosed motif can comprise helix-tum-helix (HTH), helix-loop-helix (HLH), zinc fingers, and leucine zippers. HTH and HLH are similar in that they both contain two a-helices separated by a turn (short) and a loop (long) region. Both usually bind as dimers to inverted repeats in the DNA sequence. Leucine zippers are a-helices that contain a leucine residue every seventh amino acid. This motif is found in many eukaryotic transcription factors. Zinc fingers consist of 25-30 amino acids surrounding a single zinc atom, which is coordinated by two cysteines, which are very close to short a-helices. In an aspect, a disclosed helix can contact the DNA.

[0220] In an aspect, a disclosed DNA binder can comprise a bisbenzamidine or a pyrazine.

[0221] In an aspect, a disclosed DNA binder can comprise a HuH or a dead HuH, an arginine rich motif (ARM) (e.g., a MSV ARM), an HIV enhancer-binding peptide R42, a yeast transcriptional activator GCN4, a major break region (MBR) (e.g., MBR-CC).

[0222] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise a CRISPR / Cas protein. In an aspect, a disclosed CRISPR / Cas protein can comprise a CRISPR-based endonuclease. In an aspect, a disclosed CRISPR-based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system. In an aspect, a disclosed therapeutic agent can comprise a CRISPR-based endonuclease (e.g., Cas9 or Cast 3). In an aspect, a disclosed CRISPR-based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system. In aspect, Cas9 (e.g., SpCas9 and SaCas9) are well-known to the art. In aspect, Casl3 comprises at least four known subtypes, including Casl3a (formerly C2c2), Casl3b, Cast 3 c, and Cast 3d. All known Cast 3 family members contain two HEPN domains, whichconfer RNase activity. In an aspect, Casl3 can be considered an outlier in the CRISPR world because it targets RNA, not DNA. Once it is activated by a ssRNA sequence bearing complementarity to its crRNA spacer, it unleashes a nonspecific RNase activity and destroys all nearby RNA regardless of their sequence. As disclosed herein, this property can be harnessed in vitro for precision diagnostics. Generally, Cast 3 can be found in Leptotrichia buccalis, Leptotrichia shahii, Ruminococcus flavefaciens, Bergeyella zoohelcum, Prevotella buccae, and Listeria seeligeri and can have a size of about 900 to about 1300 amino acids. In an aspect, the guide spacer length can be about 22 to about 30 nucleotides while the total guide length can be about 52 to about 66 nucleotides. In an aspect, a PAM can be 3-H for LshCasl3a, 5-D and 3- NAN or NNA for BzCasl3b, and none for RfCasl3d. In an aspect, a disclosed Casl3 can cut ssRNA.

[0223] As known to the skilled person in the cart, a Casl3d ortholog can be from a prokaryotic genome or metagenome, gut metagenome, an activated sludge metagenome, an anaerobic digester metagenome, a chicken gut metagenome, a human gut metagenome, a pig gut metagenome, a bovine gut metagenome, a sheep gut metagenome, a goat gut metagenome, a capybara gut metagenome, a primate gut metagenome, a termite gut metagenome, a fecal metagenome, a genome from the Order Clostridiales, or the Family Ruminococcaceae. In an aspect, a disclosed Cast 3d ortholog can include an Cast 3d ortholog from Ruminococcus albus, Eubacterium siraeum, a Ruminococcus flavefaciens strain XPD3002, Ruminococcus flavefaciens FD-1, uncultured Eubacterium sp TS28-c4095, uncultured Ruminococcus sp., Ruminococcus bicirculans, or Ruminococcus sp CAG57.

[0224] In an aspect, a disclosed CRISPR / Cas protein can comprise CRISPR-Cas (e.g., a small RNA-guided enzyme found uniquely in bacteriophages that can achieve programmable DNA cutting as well as genome editing).

[0225] In an aspect, a disclosed CRISPR / Cas protein can comprise a guide RNA (gRNA). In an aspect, a disclosed gRNA can comprise a specific RNA sequence that recognizes the target DNA region of interest and can direct a disclosed Cas nuclease there for editing. In an aspect, a disclosed gRNA can comprise 2 parts: (i) crispr RNA (crRNA), which is a 17-20 nucleotide sequence complementary to the targeted DNA or targeted locus, and (ii) a tracr RNA, which serves as a binding scaffold for a Cas nuclease. In an aspect, a targeted DNA or targeted locus can be a gene having one or more mutations or defects that contribute to one or more genetic diseases or disorders. In an aspect, a targeted DNA or targeted locus can be subjected to a Cas9 induced double-stranded break. In an aspect, a disclosed tracrRNA sequence can comprise nucleotides that hybridize to a minimum CRISPR repeat sequence in a cell. A minimum tracrRNA sequence and a minimum CRISPR repeat sequence may form a duplex, i.e., a base-paired double-strandedstructure. Together, the minimum tracrRNA sequence and the minimum CRISPR repeat can bind to an RNA-guided endonuclease. At least a part of the minimum tracrRNA sequence can hybridize to the minimum CRISPR repeat sequence. In an aspect, a disclosed minimum tracrRNA sequence can be at least about 30%, about 40%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or 100% complementary to the minimum CRISPR repeat sequence. For example, in an aspect, a disclosed minimum tracrRNA sequence can have a length from about 7 nucleotides to about 100 nucleotides. In an aspect, a disclsoed minimum tracrRNA sequence can be from about 7 nucleotides (nt) to about 50 nt, from about 7 nt to about 40 nt, from about 7 nt to about 30 nt, from about 7 nt to about 25 nt, from about 7 nt to about 20 nt, from about 7 nt to about 15 nt, from about 8 nt to about 40 nt, from about 8 nt to about 30 nt, from about 8 nt to about 25 nt, from about 8 nt to about 20 nt, from about 8 nt to about 15 nt, from about 15 nt to about 100 nt, from about 15 nt to about 80 nt, from about 15 nt to about 50 nt, from about 15 nt to about 40 nt, from about 15 nt to about 30 nt or from about 15 nt to about 25 nt long. In an aspect, a disclosed minimum tracrRNA sequence can be at least about 60% identical to a reference minimum tracrRNA (e.g., wild type, tracrRNA from S. pyogenes) sequence over a stretch of at least 6, 7, or 8 contiguous nucleotides. In an aspect, for example, the minimum tracrRNA sequence can be at least about 65% identical, about 70% identical, about 75% identical, about 80% identical, about 85% identical, about 90% identical, about 95% identical, about 98% identical, about 99% identical or 100% identical to a reference minimum tracrRNA sequence over a stretch of at least 6, 7, or 8 contiguous nucleotides.

[0226] In an aspect, a disclosed gRNA can target a PAM sequence at or near the targeted DNA or targeted locus. In an aspect, a spacer sequence can be designed to hybridize to a target polynucleotide that is located 5’ of a PAM of a disclosed Cas9 endonuclease. In an aspect, a disclosed spacer can match the targeted sequence or can have mismatches. In an aspect, S. pyogenes Cas9 recognizes a PAM that comprises the sequence 5’-NRG-3’, where R comprises either A or G, where N is any nucleotide and N is immediately 3’ of the target nucleic acid sequence targeted by the spacer sequence. In an aspect, S. aureus Cas9 recognizes a PAM that comprises the sequence 5'-NNGRRT-3' (where R represents A or G) an NN is immediately 3’ of the target nucleic acid sequence targeted by the spacer sequence. PAMs are known to the skilled person in the art. In an aspect, a disclosed gRNA can target a PAM sequence at or near any mutation or defect in a gene having one or more mutations or defects that contribute to one or more genetic diseases or disorders.

[0227] In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the gene expression desired. A promoter can be native or foreign and can be a natural or asynthetic sequence. By foreign, it is intended that the transcriptional initiation region is not found in the wild-type host into which the transcriptional initiation region is introduced. In an aspect, a disclosed promoter can be a promoter / enhancer. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be an endogenous promoter. In an aspect, a disclosed endogenous promoter can be an endogenous promoter / enhancer. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can generally be obtained from a non-coding region upstream of a transcription initiation site of a gene of interest. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can be used for constitutive and efficient expression of a disclosed gene. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be a CMV promoter or a CMV promoter / enhancer. CMV promoters and CMV promoters / enhancers are well known to the art. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be any eukaryotic RNA polymerase II promoter.

[0228] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an oligonucleotide agent or an oligonucleotide therapeutic agent. In an aspect, a disclosed oligonucleotide agent or an oligonucleotide therapeutic agent can be a single-stranded or double-stranded DNA, iRNA, shRNA, siRNA, mRNA, non-coding RNA (ncRNA), an antisense molecule, miRNA, a morpholino, a peptide-nucleic acid (PNA), or an analog or conjugate thereof. In an aspect, a disclosed cargo can be an ASO or an RNAi.

[0229] In an aspect, a disclosed cargo can comprise ribosomal RNA (rRNA), transfer RNA (tRNA), heterogeneous nuclear RNA (hnRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), Piwi-interacting RNA (piRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), singe guide RNA (sgRNA), non-coding RNA (ncRNA), long non-coding RNA (IncRNA), 7SL, Xist, short enhancer RNA (eRNA), circular RNA, intergenic RNA, or any combination thereof. In an aspect, a disclosed RNA can comprise IncRNA, siRNA, shRNA, sgRNA, circular RNA, snoRNA, miRNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise a functional non-coding RNA element.

[0230] In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed RNA can comprise ribosomal RNA (rRNA), transfer RNA (tRNA), heterogeneous nuclear RNA (hnRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), Piwi-interacting RNA (piRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), singe guide RNA (sgRNA), non-coding RNA (ncRNA), long non-coding RNA (IncRNA), 7SL, Xist, short enhancer RNA (eRNA), circular RNA, intergenic RNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise IncRNA, siRNA, shRNA, sgRNA, circularRNA, snoRNA, miRNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise a functional non-coding RNA element.

[0231] Disclosed herein are pharmaceutical formulations comprising one or more disclosed AAV particles and one or more carriers, diluents, and / or excipients. Disclosed herein are pharmaceutical formulations comprising one or more disclosed AAV-like particles and one or more carriers, diluents, and / or excipients.

[0232] Disclosed herein are pharmaceutical formulations comprising one or more disclosed AAV particles and one or more pharmaceutically acceptable carriers. Disclosed herein are pharmaceutical formulations comprising one or more disclosed AAV-like particles and one or more pharmaceutically acceptable carriers.7. Nanocontainers

[0233] Disclosed herein is a nanocontainer comprising an AAV particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a nanocontainer comprising an AAV-like particle comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a nanocontainer comprising a plurality of AAV particles comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a nanocontainer comprising a plurality AAV-like particles comprising a disclosed J peptide fused to a disclosed cargo. Disclosed herein is a nanocontainer comprising an AAV particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a nanocontainer comprising an AAV-like particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a nanocontainer comprising a plurality of AAV particles comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a nanocontainer comprising a plurality AAV-like particle comprising a disclosed cargo fused to a disclosed J peptide. Disclosed herein is a nanocontainer comprising an AAV particle comprising a disclosed cargo or disclosed cargos. Disclosed herein is a nanocontainer comprising an AAV- like particle comprising a disclosed cargo or disclosed cargos. Disclosed herein is a nanocontainer comprising a plurality of AAV particles comprising a disclosed cargo or disclosed cargos. Disclosed herein is a nanocontainer comprising a plurality AAV-like particles comprising a disclosed cargo or disclosed cargos.

[0234] In an aspect, a disclosed nanocontainer comprising an AAV-like particle can be non- infectious and unable to replicate due to the absence of a viral genome. In an aspect of a disclosed nanocontainer, a plurality of disclosed AAV-like particles can be non-infectious and unable to replicate due to the absence of a viral genome.

[0235] In an aspect, a disclosed nanocontainer comprising a disclosed AAV particle and / or a disclosed AAV-like particle comprising a disclosed cargo can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme. In an aspect of adisclosed nanocontainer, a disclosed AAV particle and / or a disclosed AAV-like particle comprising a disclosed cargo can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, restoring one or more aspect of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation comprises restoring the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme. In an aspect of a disclosed nanocontainer, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell types; (ii) normalizing aspects of the autophagy pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or preserving organelle functionality and / or structural integrity; (v) correcting enzyme dysregulation; (vi) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the multi -systemic manifestations of a genetic disease or disorder; (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of a genetic disease or disorder, or (viii) any combination thereof.

[0236] In an aspect of a disclosed nanocontainer, restoring the activity and / or functionality of a missing, deficient, and / or mutant protein or enzyme can comprise a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10- 20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a control level or a reference level (e.g., determined, for example, using one or more subjects not having a missing, deficient, and / or mutant protein or enzyme). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity, and / or functionality is similar to that of a wild-type or control level.

[0237] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a protein or a therapeutic protein. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an enzyme or a therapeutic enzyme. In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a therapeutic cargo. In an aspect, a disclosed therapeutic cargo can comprise a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject. In an aspect of a disclosed nanocontainer, a disclosed therapeutic cargo can replace and / or supplement a protein or an enzyme that is missing, deficient, and / or mutated in cell and / or in a subject.

[0238] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a non-DNA target. In an aspect of a disclosed AAV particle and / or a disclosed AAV-like particle, a disclosed cargo can comprise an RNA binder. In an aspect of a disclosed nanocontainer, a disclosed RNA binder can comprise a small molecule that binds to a target RNA. In an aspect of a disclosed nanocontainer, a disclosed RNA binder can comprise a macrolide, an alkaloid, an aminoglycoside, an anthracene, an oxazolidinone, a ribocil, an SMN2 ligand, a tetracycline, or a triptycene.

[0239] In an aspect of a disclosed nanocontainer, a disclosed small molecule that binds to a target RNA is an anthracene, a kanamycin, a linezolid, a neomycin, a paromomycin, a pleuromutilin, a ribocil, a tedizolid, or a triptycene.

[0240] In an aspect of a disclosed nanocontainer, a disclosed RNA binder can comprise a CPMV arginine rich motif (ARM), a STNV arginine rich motif (ARM), LambdaN, or a MS2. In an aspect, a disclosed RNA binder can comprise roseoflavin.

[0241] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a molecular glue. In an aspect, a disclosed MG can comprise cyclosporin, FK506, thalidomide, lenalidomide, or any combination thereof.

[0242] In an aspect, a disclosed molecular glue (MG) can be a small molecule that possess intrinsic property to induce interaction between two proteins. In an aspect, a disclosed MG can promote the dimerization or colocalization between two unrelated proteins via formation of a ternary complex. In an aspect, these induced interactions can be between a target protein and a ubiquitin ligase, and can result in its proteasomal degradation or between two unrelated proteins and a perturbation of signaling cascades.

[0243] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise an epigenetic modulator for activation or repression. In an aspect, a disclosed epigenetic modulator can comprise a polypeptide having some type of activation activity or repression activity. In an aspect, a disclosed epigenetic modulator can comprise a DNA methyltransferase (DNMT) inhibitor, a histone methyltransferase, a demethylase, an acetyltransferase, a deacetylase, a bromodomain and extra-terminal domain (BET) inhibitor, any fusion thereof, or any combination thereof.

[0244] In an aspect of a disclosed nanocontainer, a disclosed epigenetic modulator can comprise a polypeptide having transcription activation activity, transcription repression activity, transcription release factor activity, histone modification activity, nucleic acid association activity, methyltransferase activity, demethylase activity, acetyltransferase activity, deacetylase activity, or any combination thereof.

[0245] In an aspect of a disclosed nanocontainer, a disclosed encoded polypeptide can comprise HP la, HP lb, MBD1, MBD2, Kriippel -Associated Box (KRAB), NIPP1, the Transcription Repression Domain (TRD) of Methyl-CpG Binding Protein 2 (MeCP2), DNMT3 A, any fusionthereof, or any combination thereof. In an aspect, a disclosed fusion or disclosed fusion protein can comprise HP 1 a-HP 1 a, HP 1 a-HP lb, HP 1 a-MBD 1 , HP 1 a-MBD2, HP 1 a-KRAB, HP 1 a-NIPP 1 , HPla-MeCP2, HPla-DNMT3A, HPlb-HPlb, HPlb-HPla, HPlb-MBDl, HPlb-MBD2, HPlb- KRAB, HPlb-NIPPl, HPlb-MeCP2, HPlb-DNMT3A, MBD1-MBD1, MBDl-HPla, MBD1- HPlb, MBD1-MBD2, MBD1-KRAB, MBD1-NIPP1, MBDl-MeCP2, MBD1-DNMT3A, MBD2-MBD2, MBD2-HPla, MBD2-HPlb, MBD2-MBD1, MBD2-KRAB, MBD2-NIPP1, MBD2-MeCP2, MBD2-DNMT3A, KRAB-KRAB, KRAB-HPla, KRAB-HPlb, KRAB-MBD1, KRAB-MBD2, KRAB-NIPP1, KRAB-MeCP2, KRAB-DNMT3A, NIPPI-NIPPI, NIPPl-HPla, NIPPl-HPlb, NIPP1-MBD1, NIPP1-MBD2, NIPP1-KRAB, NIPPl-MeCP2, NIPP1-DNMT3A, MeCP2-MeCP2, MeCP2-HPla, MeCP2-HPlb, MeCP2-MBDl, MeCP2-MBD2, MeCP2-KRAB, MeCP2-NLPPl, MeCP2-DNMT3A, DNMT3A-DNMT3A, DNMT3A-HPla, DNMT3A-HPlb, DNMT3A-MBD1, DNMT3A-MBD2, DNMT3 A-KRAB, DNMT3A-NIPP1, or DNMT3A- MeCP2.

[0246] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a transcription factor or a binder thereof. In an aspect, exemplary transcription factors can include, but are not limited to, nuclear factor KB (NF-KB) (e.g., 50, p52, c-Rel, p65 (Rel-A), and Rel-B), activating protein 1 (AP-1), signal transducer and activator of transcription factor (STAT)-3, and members of the CCAAT / enhancer-binding protein (CZEBP) family of transcription factors (e.g., CZEBPP and C / EBP5).

[0247] As known in the art, transcription factors (TFs) recognize specific DNA sequences to control chromatin and transcription, forming a complex system that guides expression of the genome.

[0248] As reviewed by Lambert SA et al. (2018), there are over 1600 likely human TFs and binding motifs for two-thirds of them. TFs cannot be understood functionally without accompanying detailed knowledge of the DNA sequences they bind. TF DNA-binding specificities are frequently summarized as “motifs” - models representing the set of related, short sequences preferred by a given TF, which can be used to scan longer sequences (e.g., promoters) to identify potential binding sites. Determining a DNA-binding motif is often the first step toward detailed examination of the function of a TF because identification of potential binding sites provides a gateway to further analyses.

[0249] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise an inhibitor of a cellular degradation process.

[0250] In an aspect of a disclosed nanocontainer, a disclosed inhibitor of a cellular degradation process can comprise a protease blocker or inhibitor and / or a proteosome blocker or inhibitor. In an aspect, a disclosed protease and / or proteasome blocker or inhibitor can comprise Pepstatin A(can Inhibit aspartic acid proteases), Leupeptin (can inhibit serine and cysteine proteases), EDTA (can inhibit metalloproteases), PMSF (can inhibit serine proteases), Bortezomib (e.g., a proteasome inhibitor that is a slowly reversible inhibitor), Carfilzomib (e.g., a proteasome inhibitor that is an irreversible inhibitor), Delanzomib, Ixazomib (e.g., a proteasome inhibitor that is a reversible inhibitor), or any combination thereof. In an aspect, a disclosed inhibitor of a cellular degradation process can comprise an autophagy inhibitor. In an aspect, a disclosed autophagy inhibitor can comprise E64d (e.g., a membrane-permeable inhibitor of cathepsins B, H, and L), Pepstatin A (can inhibit cathepsins D and E), Baf Al (can inhibit vacuolar H+ ATPase), Monensin (can mediate the exchange of protons for potassium or sodium), 3 -MA (can prevent the starvation-induced colocalization of mitochondrial and lysosomal markers), or any combination thereof.

[0251] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a DNA binder. In an aspect, a disclosed DNA binder can be a small molecule (can bind to DNA through intercalation or minor groove binding), a DNA-binding protein (can bind to DNA through domains such as helix-turn-helix, zinc finger, and homeodomains), a bisbenzamidine (can bind to the minor groove of DNA), a photosensitive metal complex (can bind to DNA duplexes), or any combination thereof.

[0252] In an aspect of a disclosed nanocontainer, a disclosed DNA binder can alter the stability and functionality of DNA, can be used to program reaction pathways of dynamic DNA reactions, can be used to trigger site-selective, DNA-promoted biochemical and physicochemical processes, or any combination thereof. In an aspect, a disclosed DNA binder can be used to accelerate the discovery and characterization of bioactive small molecules, to develop high-throughput screening assays for discovering DNA binders, or any combination thereof.

[0253] In an aspect of a disclosed nanocontainer, a disclosed DNA binder can bind to DNA, thereby altering the structure and / or function of DNA. In an aspect, a disclosed DNA binding protein can comprise a polypeptide motif that enables the binding to the major groove of DNA. In an aspect, a disclosed motif can comprise helix-tum-helix (HTH), helix-loop-helix (HLH), zinc fingers, and leucine zippers. HTH and HLH are similar in that they both contain two a-helices separated by a turn (short) and a loop (long) region. Both usually bind as dimers to inverted repeats in the DNA sequence. Leucine zippers are a-helices that contain a leucine residue every seventh amino acid. This motif is found in many eukaryotic transcription factors. Zinc fingers consist of 25-30 amino acids surrounding a single zinc atom, which is coordinated by two cysteines, which are very close to short a-helices. In an aspect, a disclosed helix can contact the DNA.

[0254] In an aspect of a disclosed nanocontainer, a disclosed DNA binder can comprise a bisbenzamidine or a pyrazine.

[0255] In an aspect of a disclosed nanocontainer, a disclosed DNA binder can comprise a HuH or a dead HuH, an arginine rich motif (ARM) (e.g., a MSV ARM), an HIV enhancer-binding peptide R42, a yeast transcriptional activator GCN4, a major break region (MBR) (e.g., MBR-CC).

[0256] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise a CRISPR / Cas protein. In an aspect, a disclosed CRISPR / Cas protein can comprise a CRISPR-based endonuclease. In an aspect, a disclosed CRISPR-based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system. In an aspect, a disclosed therapeutic agent can comprise a CRISPR-based endonuclease (e.g., Cas9 or Cast 3). In an aspect, a disclosed CRISPR- based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system. In aspect, Cas9 (e.g., SpCas9 and SaCas9) are well-known to the art. In aspect, Casl3 comprises at least four known subtypes, including Cast 3a (formerly C2c2), Cast 3b, Cast 3 c, and Cast 3d. All known Cast 3 family members contain two HEPN domains, which confer RNase activity. In an aspect, Cas 13 can be considered an outlier in the CRISPR world because it targets RNA, not DNA. Once it is activated by a ssRNA sequence bearing complementarity to its crRNA spacer, it unleashes a nonspecific RNase activity and destroys all nearby RNA regardless of their sequence. As disclosed herein, this property can be harnessed in vitro for precision diagnostics. Generally, Cas 13 can be found in Leptotrichia buccalis, Leptotrichia shahii, Ruminococcus flavefaciens, Bergeyella zoohelcum, Prevotella buccae, and Listeria seeligeri and can have a size of about 900 to about 1300 amino acids. In an aspect, the guide spacer length can be about 22 to about 30 nucleotides while the total guide length can be about 52 to about 66 nucleotides. In an aspect, a PAM can be 3-H for LshCasl3a, 5-D and 3 -NAN orNNA for BzCasl3b, and none for RfCasl3d. In an aspect, a disclosed Cas 13 can cut ssRNA.

[0257] As known to the skilled person in the cart, a Cas 13d ortholog can be from a prokaryotic genome or metagenome, gut metagenome, an activated sludge metagenome, an anaerobic digester metagenome, a chicken gut metagenome, a human gut metagenome, a pig gut metagenome, a bovine gut metagenome, a sheep gut metagenome, a goat gut metagenome, a capybara gut metagenome, a primate gut metagenome, a termite gut metagenome, a fecal metagenome, a genome from the Order Clostridiales, or the Family Ruminococcaceae. In an aspect, a disclosed Cas 13d ortholog can include an Cast 3d ortholog from Ruminococcus albus, Eubacterium siraeum, a Ruminococcus flavefaciens strain XPD3002, Ruminococcus flavefaciens FD-1, uncultured Eubacterium sp TS28-c4095, uncultured Ruminococcus sp., Ruminococcus bicirculans, or Ruminococcus sp CAG57.

[0258] In an aspect of a disclosed nanocontainer, a disclosed CRISPR / Cas protein can comprise CRISPR-Cas (e.g., a small RNA-guided enzyme found uniquely in bacteriophages that can achieve programmable DNA cutting as well as genome editing).

[0259] In an aspect of a disclosed nanocontainer, a disclosed CRISPR / Cas protein can comprise a guide RNA (gRNA). In an aspect, a disclosed gRNA can comprise a specific RNA sequence that recognizes the target DNA region of interest and can direct a disclosed Cas nuclease there for editing. In an aspect, a disclosed gRNA can comprise 2 parts: (i) crispr RNA (crRNA), which is a 17-20 nucleotide sequence complementary to the targeted DNA or targeted locus, and (ii) a tracr RNA, which serves as a binding scaffold for a Cas nuclease. In an aspect, a targeted DNA or targeted locus can be a gene having one or more mutations or defects that contribute to one or more genetic diseases or disorders. In an aspect, a targeted DNA or targeted locus can be subj ected to a Cas9 induced double-stranded break. In an aspect, a disclosed tracrRNA sequence can comprise nucleotides that hybridize to a minimum CRISPR repeat sequence in a cell. A minimum tracrRNA sequence and a minimum CRISPR repeat sequence may form a duplex, i.e., a basepaired double-stranded structure. Together, the minimum tracrRNA sequence and the minimum CRISPR repeat can bind to an RNA-guided endonuclease. At least a part of the minimum tracrRNA sequence can hybridize to the minimum CRISPR repeat sequence. In an aspect, a disclosed minimum tracrRNA sequence can be at least about 30%, about 40%, about 50%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or 100% complementary to the minimum CRISPR repeat sequence. For example, in an aspect, a disclosed minimum tracrRNA sequence can have a length from about 7 nucleotides to about 100 nucleotides. In an aspect, a disclosed minimum tracrRNA sequence can be from about 7 nucleotides (nt) to about 50 nt, from about 7 nt to about 40 nt, from about 7 nt to about 30 nt, from about 7 nt to about 25 nt, from about 7 nt to about 20 nt, from about 7 nt to about 15 nt, from about 8 nt to about 40 nt, from about 8 nt to about 30 nt, from about 8 nt to about 25 nt, from about 8 nt to about 20 nt, from about 8 nt to about 15 nt, from about 15 nt to about 100 nt, from about 15 nt to about 80 nt, from about 15 nt to about 50 nt, from about 15 nt to about 40 nt, from about 15 nt to about 30 nt or from about 15 nt to about 25 nt long. In an aspect, a disclosed minimum tracrRNA sequence can be at least about 60% identical to a reference minimum tracrRNA (e.g., wild type, tracrRNA from S. pyogenes) sequence over a stretch of at least 6, 7, or 8 contiguous nucleotides. In an aspect, for example, the minimum tracrRNA sequence can be at least about 65% identical, about 70% identical, about 75% identical, about 80% identical, about 85% identical, about 90% identical, about 95% identical, about 98% identical, about 99% identical or 100% identical to a reference minimum tracrRNA sequence over a stretch of at least 6, 7, or 8 contiguous nucleotides.

[0260] In an aspect of a disclosed nanocontainer, a disclosed gRNA can target a PAM sequence at or near the targeted DNA or targeted locus. In an aspect, a spacer sequence can be designed to hybridize to a target polynucleotide that is located 5’ of a PAM of a disclosed Cas9 endonuclease. In an aspect, a disclosed spacer can match the targeted sequence or can have mismatches. In an aspect, S. pyogenes Cas9 recognizes a PAM that comprises the sequence 5’-NRG-3’, where R comprises either A or G, where N is any nucleotide and N is immediately 3’ of the target nucleic acid sequence targeted by the spacer sequence. In an aspect, S. aureus Cas9 recognizes a PAM that comprises the sequence 5'-NNGRRT-3' (where R represents A or G) an NN is immediately 3’ of the target nucleic acid sequence targeted by the spacer sequence. PAMs are known to the skilled person in the art. In an aspect, a disclosed gRNA can target a PAM sequence at or near any mutation or defect in a gene having one or more mutations or defects that contribute to one or more genetic diseases or disorders.

[0261] In an aspect of a disclosed nanocontainer, a disclosed promoter for the one or more disclosed guide RNA sequences can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the gene expression desired. A promoter can be native or foreign and can be a natural or a synthetic sequence. By foreign, it is intended that the transcriptional initiation region is not found in the wild-type host into which the transcriptional initiation region is introduced. In an aspect, a disclosed promoter can be a promoter / enhancer. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be an endogenous promoter. In an aspect, a disclosed endogenous promoter can be an endogenous promoter / enhancer. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can generally be obtained from a non-coding region upstream of a transcription initiation site of a gene of interest. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can be used for constitutive and efficient expression of a disclosed gene. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be a CMV promoter or a CMV promoter / enhancer. CMV promoters and CMV promoters / enhancers are well known to the art. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be any eukaryotic RNA polymerase II promoter.

[0262] In an aspect of a disclosed nanocontainer a disclosed cargo can comprise an oligonucleotide agent or an oligonucleotide therapeutic agent. In an aspect, a disclosed oligonucleotide agent or an oligonucleotide therapeutic agent can be a single-stranded or doublestranded DNA, iRNA, shRNA, siRNA, mRNA, non-coding RNA (ncRNA), an antisense molecule, miRNA, a morpholino, a peptide-nucleic acid (PNA), or an analog or conjugate thereof. In an aspect, a disclosed cargo can be an ASO or an RNAi.

[0263] In an aspect of a disclosed nanocontainer, a disclosed cargo can comprise ribosomal RNA (rRNA), transfer RNA (tRNA), heterogeneous nuclear RNA (hnRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), Pi wi -interacting RNA (piRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), singe guide RNA (sgRNA), non-coding RNA (ncRNA), long non-coding RNA (IncRNA), 7SL, Xist, short enhancer RNA (eRNA), circular RNA, intergenic RNA, or any combination thereof. In an aspect, a disclosed RNA can comprise IncRNA, siRNA, shRNA, sgRNA, circular RNA, snoRNA, miRNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise a functional non-coding RNA element.

[0264] In an aspect of a disclosed nanocontainer, a disclosed RNA can comprise ribosomal RNA (rRNA), transfer RNA (tRNA), heterogeneous nuclear RNA (hnRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), Piwi-interacting RNA (piRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), singe guide RNA (sgRNA), non-coding RNA (ncRNA), long non-coding RNA (IncRNA), 7SL, Xist, short enhancer RNA (eRNA), circular RNA, intergenic RNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise IncRNA, siRNA, shRNA, sgRNA, circular RNA, snoRNA, miRNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise a functional non-coding RNA element.

[0265] Disclosed herein are pharmaceutical formulations comprising a nanocontainer comprising one or more disclosed AAV particles and one or more carriers, diluents, and / or excipients. Disclosed herein are pharmaceutical formulations comprising a nanocontainer comprising one or more disclosed AAV-like particles and one or more carriers, diluents, and / or excipients.Disclosed herein are pharmaceutical formulations comprising a nanocontainer comprising one or more disclosed AAV particles and one or more pharmaceutically acceptable carriers. Disclosed herein are pharmaceutical formulations comprising a nanocontainer comprising one or more disclosed AAV-like particles and one or more pharmaceutically acceptable carriers.8. Vectors

[0266] Disclosed herein is a vector comprising a disclosed nucleic acid molecule. Disclosed herein is a vector comprising one or more disclosed nucleic acid molecules. Disclosed herein is a vector comprising a disclosed nucleic acid molecule encoding a disclosed J peptide fused to or linked with a disclosed cargo. Disclosed herein is a vector comprising one or more disclosed nucleic acid molecules. In an aspect, a disclosed nucleic acid sequence can have a coding sequence that is less than about 4.5 kilobases.

[0267] In an aspect, a disclosed vector can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, a disclosed vector canrestore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme.

[0268] In an aspect, a disclosed vector can be a viral vector or a non-viral vector. In an aspect, a disclosed non-viral vector can be a polymer-based vector, a peptide-based vector, a lipid nanoparticle, a solid lipid nanoparticle, or a cationic lipid-based vector. In an aspect, a disclosed vector can comprise exosomes, extracellular vesicles, and virus like particles. In an aspect, a disclosed viral vector can be an adenovirus vector, an AAV vector, a herpes simplex virus vector, a retrovirus vector, a lentivirus vector, and alphavirus vector, a Flavivirus vector, a rhabdovirus vector, a measles virus vector, a Newcastle disease viral vector, a poxvirus vector, or a picomavirus vector.

[0269] In an aspect, a disclosed viral vector can be an adeno-associated virus (AAV) vector In an aspect, a disclosed AAV vector can include naturally isolated serotypes including, but not limited to, AAV1, AAV2, AAV3 (including 3a and 3b), AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAV9, AAV10, AAVrhlO, AAV11, AAV12, AAV13, AAVrh39, AAVrh43, AAVcy.7 as well as bovine AAV, caprine AAV, canine AAV, equine AAV, ovine AAV, avian AAV, primate AAV, non-primate AAV, and any other virus classified by the International Committee on Taxonomy of Viruses (ICTV) as an AAV. In an aspect, an AAV capsid can be a chimera either created by capsid evolution or by rational capsid engineering from a naturally isolated AAV variants to capture desirable serotype features such as enhanced or specific tissue tropism and / or a host immune response escape. Naturally isolated AAV variants include, but not limited to, AAV-DJ, AAV-HAE1, AAV-HAE2, AAVM41, AAV-1829, AAV2 Y / F, AAV2 T / V, AAV2i8, AAV2.5, AAV9.45, AAV9.61, AAV-B1, AAV-AS, AAV9.45A-String (e.g., AAV9.45-AS), AAV9.45Angiopep, AAV9.47-Angiopep, and AAV9.47-AS, AAV-PHP.B, AAV-PHP.eB, AAV-PHP.S, AAV-F, AAVcc.47, and AAVcc.81. In an aspect, a disclosed AAV vector can be AAV-Rh74 or a related variant (e.g., capsid variants like RHM4-1). In an aspect, a disclosed AAV vector can be AAV8. In an aspect, a disclosed AAV vector can be AAVhum.8. In an aspect, a disclosed AAV vector can be a self-complementary AAV as disclosed herein.9. Cells

[0270] Disclosed herein are cells comprising a disclosed nucleic acid molecule, a disclosed vector, and / or a disclosed plasmid. Disclosed herein are cells subjected to triple transfection using a disclosed plasmid comprising a nucleic acid sequence encoding a disclosed J peptide. Disclosed herein are cells subjected to triple transfection using a disclosed plasmid comprising a nucleic acid sequence encoding a disclosed J peptide fused to a disclosed cargo. Disclosed herein are cells transduced by one or more disclosed viral vectors. Disclosed herein are cells transfected with one or more disclosed nucleic acid molecules. In an aspect, a disclosed cell has been transfected withone or more nucleic acid sequences having the sequence set forth in any of SEQ ID NO:49-SEQ ID NO:98. In an aspect, a disclosed cell has been transfected with one or more nucleic acid sequences having the sequence set forth in any of SEQ ID NO:99-SEQ ID NO: 126.

[0271] Techniques to achieve transfection and transduction are known to the art and using transfected or transduced cells are known to the art. In an aspect, disclosed herein are human immortalized cells lines transduced by one or more disclosed viral vectors or transfected with one or more disclosed nucleic acids or disclosed plasmids. In an aspect, disclosed herein are human immortalized cells lines contacted with one or more disclosed pharmaceutical formulations. Disclosed herein are cells obtained for a subject treated with one or more disclosed nucleic acid molecule, one or more disclosed vectors, one or more disclosed plasmids, or one or more disclosed pharmaceutical formulations. Disclosed herein are cells that generate one or more disclosed AAV particles or one or more disclosed AAV-like particles.10. Kits

[0272] Disclosed herein is a kit comprising one or more of a disclosed J peptide, a disclosed nucleic acid molecule, a disclosed plasmid, a disclosed vector, a disclosed pharmaceutical formulation, or any combination thereof. Disclosed herein is a kit comprising one or more disclosed J peptides, one or more disclosed nucleic acid molecules, one or more disclosed vectors, one or more disclosed pharmaceutical formulations, or any combination thereof. In an aspect, a kit can comprise a disclosed J peptide, a disclosed plasmid, a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, a disclosed RNA therapeutic, or a combination thereof, and one or more agents. “Agents” and “Therapeutic Agents” are known to the art and are described supra. In an aspect, the one or more agents can treat, prevent, inhibit, and / or ameliorate one or more comorbidities in a subject. In an aspect, one or more active agents can treat, inhibit, prevent, and / or ameliorate cellular and / or metabolic complications related to a missing, deficient, and / or mutant protein or enzyme.

[0273] Disclosed here is a kit comprising one or more disclosed AAV particles and / or one or more AAV-like particles.

[0274] In an aspect, a disclosed kit can comprise at least two components constituting the kit. Together, the components constitute a functional unit for a given purpose (such as, for example, treating a subject diagnosed with or suspected of having a disease or disorder). Individual member components may be physically packaged together or separately. For example, a kit comprising an instruction for using the kit may or may not physically include the instruction with other individual member components. Instead, the instruction can be supplied as a separate member component, either in a paper form or an electronic form which may be supplied on computer readable memory device or downloaded from an internet website, or as recorded presentation. Inan aspect, a kit for use in a disclosed method can comprise one or more containers holding disclosed J peptide, a disclosed plasmid, a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, or a combination thereof, and a label or package insert with instructions for use. In an aspect, suitable containers include, for example, bottles, vials, syringes, blister pack, etc. The containers can be formed from a variety of materials such as glass or plastic. The container can hold a disclosed J peptide, a disclosed plasmid, a disclosed cargo, a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, or a combination thereof, and can have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The label or package insert can indicate that a disclosed J pepide, a disclosed plasmid, a disclosed cargo, a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, a disclosed RNA therapeutic, or a combination thereof can be used for treating, preventing, inhibiting, and / or ameliorating a disease or disorder or complications and / or symptoms associated with a disease or disorder. A kit can comprise additional components necessary for administration such as, for example, other buffers, diluents, filters, needles, and syringes.

[0275] Disclosed herein is a kit comprising one or more agents and reagents that are necessary to create disclosed AAV particles or disclosed AAV-like particles using a triple transfection protocol or a modified triple transfection protocol. In an aspect, a disclosed kit can comprise a disclosed J peptide, a disclosed plasmid, a disclosed cargo, a disclosed producer cell, or any combination thereof. In an aspect, transfection of pTrans:pCis:pHelper are in a 1 :0.6: 1.2 ratio. In an aspect, a disclosed kit can be used to load a fused cargo into one or more AAV particles or one or more AAV-like particles. In an aspect, a disclosed kit can be used to generate AAV particles and / or AAV-like particles comprising a desired cargo fused to a disclosed J peptide. In an aspect, a disclosed kit can be used to treat a subject or slow the progression of a disease or disorder in a subject.C. Methods1. Methods of Loading and / or Generating an AAV Particle or AAV-Like Particle

[0276] Disclosed herein is a method of loading a disclosed cargo into an AAV particle, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0277] Disclosed herein is a method of loading a cargo into an AAV particle, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfectioncomprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0278] Disclosed herein is a method of loading a cargo into an AAV particle, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0279] Disclosed herein is a method of loading a disclosed cargo into an AAV-like particle, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0280] Disclosed herein is a method of loading a cargo into an AAV-like particle, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0281] Disclosed herein is a method of loading a cargo into an AAV-like particle, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0282] Disclosed herein is a method of generating an AAV particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0283] Disclosed herein is a method of generating an AAV particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0284] Disclosed herein is a method of generating an AAV particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0285] Disclosed herein is a method of generating an AAV-like particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed peptide linked to and / or fused with a disclosed cargo.

[0286] Disclosed herein is a method of generating an AAV-like particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises a plasmid comprising a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo.

[0287] Disclosed herein is a method of generating an AAV-like particle loaded with a disclosed cargo, the method comprising: using triple transfection to produce AAV-like particles, wherein the triple transfection comprises (i) a helper plasmid (pHelper); (ii) a trans-complementing plasmid (pTrans); and (iii) a cis plasmid (pCis).

[0288] In an aspect, a disclosed helper plasmid or a disclosed pHelper can comprise or can deliver one or more adenovirus helper genes. In an aspect, a disclosed trans-complementing plasmid or a disclosed pTrans can comprise or can express AAV Cap genes and optionally AAV Rep genes. In an aspect, a disclosed trans-complementing plasmid or a disclosed pTrans can comprise or can express an AAV Cap gene. In an aspect, a disclosed trans-complementing plasmid or a disclosed pTrans can comprise or can express a fragment of the AAV Cap gene by a non-endogenous promoter. In an aspect, a disclosed trans-complementing plasmid or a disclosed pTrans can comprise or can express AAV Cap gene by a non-endogenous promoter starting from the P40 transcription start site that retains a non-functional fragment of Rep.

[0289] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs).

[0290] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs).

[0291] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:15.

[0292] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0293] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11.

[0294] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 11.

[0295] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0296] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence for a disclosed J peptide linked to and / or fused with a disclosed cargo, wherein the nucleic acid sequence is flanked by AAV inverted terminal repeats (ITRs), wherein the disclosed encoded J peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO:29.

[0297] Triple transfection and methods to effect triple transfection are known to the art. Kits and components for triple transfection are commercially available. As known to the art, for example, AAV requires a helper plasmid containing genes from adenovirus that mediate AAV replication (i.e., E4, E2a and VA). In an aspect of a disclosed method, triple transfection can comprise using a pHelper plasmid, a plasmid comprising Rep / Cap, and a transfer plasmid comprising a disclosed J peptide or disclosed peptide fused to a disclosed cargo. In an aspect, generating AAV can occur in suspension cells. In an aspect of a disclosed method, suspension cells can be incubated. In an aspect, suspension cells (e.g., HEK cells, HEKG2 cells, etc.) and methods of incubating suspension cells. In an aspect, a disclosed method can comprise incubating suspension cells.

[0298] In an aspect, disclosed triple transfection method can comprise employing a 4thplasmid. In an aspect, a disclosed 4thplasmid can comprise a desired nucleic acid sequence. In an aspect, this system can comprise pHelper, pRep / Cap, pITR, and pCis. In an aspect, a disclosed transfection ratio for pHelper:pRep / Cap:pITR:pC can be 1.2: 1 :0.6:0.6. In an aspect the pITRencodes a therapeutic gene flanked by ITRs for packaging into the AAV capsid and pCis encodes a functional peptide or protein fused to J peptide for co-packaging with the genome. In an aspect, the functional peptide or protein can control or regulate expression from the therapeutic gene copackaged into the AAV capsid. In an aspect, the functional peptide or protein can be an inhibitor of host-cell factors that restrict expression from the therapeutic gene co-packaged into the AAV capsid.

[0299] In an aspect, a disclosed method can generate AAV particles or AAV-like particles that comprise an RNA cargo.

[0300] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid.

[0301] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a “J peptide”. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid.

[0302] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta Gstrand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 15.

[0303] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0304] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV), wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0305] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0306] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV), wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 to SEQ ID NO: 15.

[0307] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis cancomprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11.

[0308] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO: 11.

[0309] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis cancomprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

[0310] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of an adeno-associated virus (AAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the luminal surface of a recombinant adeno-associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide targeting the beta G strand of the capsid luminal surface of a recombinant adeno- associated virus (rAAV) capsid, wherein the encoded peptide comprises a sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

[0311] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%,or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:98.

[0312] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:64. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:60. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising the nucleic acid sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or more than at least 95% identity to the sequence set forth in any one of SEQ ID NO:50 - SEQ ID NO:78.

[0313] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide fused to a linker. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a J peptide fused to a linker.

[0314] In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise a sequence having about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126 and can be linked to a disclosed cargo. In an aspect, a disclosed cis plasmid or a disclosed pCis can comprise a cassette comprising a nucleic acid sequence encoding a peptide fused to a linker or a disclosed J peptide fused to a linker can comprise a sequence having about 95%, about 96%, about 97%,about 98%, about 99%, or more than about 99% identity to the sequence set forth in any one of SEQ ID NO:99 - SEQ ID NO: 126 and can be linked to a disclosed cargo.

[0315] Disclosed herein is a method of loading a cargo into an AAV particle, the method comprising: using a modified transfection protocol to produce AAV particles, wherein the modified transfection protocol comprises (i) a helper plasmid (pHelper); and (ii) a cis plasmid (pCis) comprising a nucleic acid sequence encoding a disclosed J peptide.

[0316] In an aspect, a disclosed modified transfection protocol does not include a functional Rep. In an aspect, a disclosed modified transfection protocol does not employ a functional Rep but employs 3 plasmids.

[0317] In an aspect of a disclosed cis plasmid or a disclosed pCis, a disclosed cassette can comprise one or more regulatory elements. In an aspect, one or more disclosed regulatory elements can comprise an internal ribosomal entry site (IRES), a promoter, an enhancer, a promoter / enhancer, one or more inverted terminal repeats (ITRs), a poly adenylation sequence, a transcription termination signal, any combination thereof, or a combination of all disclosed regulatory elements. ITRs are known to the skilled person. ITRs can be AAV ITRs, such as, for example, ITRs from AAV1, AAV2, AAV3a and AAV3b, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV 10, AAV11, AAV 12, AAV13, or any other naturally occurring AAV. In an aspect, ITRs can be modified. In an aspect, ITRs can be from a recombinant AAV.

[0318] In an aspect, a disclosed promoter can be operably linked to (i) a nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide, or (ii) a nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide linked to a disclosed cargo.

[0319] In an aspect, a disclosed promoter and / or enhancer can be positioned 5’ (upstream) or 3’ (downstream) of a disclosed nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide under its control. As is known in the art, the distance between the placement of a promoter and / or enhancer from a disclosed nucleic acid sequence encoding a disclosed peptide or a disclosed J peptide variation can be accommodated without loss of function.

[0320] Disclosed herein is an AAV particle comprising a disclosed J peptide fused to a disclosed cargo made by a disclosed method. Disclosed herein is an AAV-like particle comprising a disclosed J peptide fused to a disclosed cargo made by a disclosed method.

[0321] In an aspect, a disclosed method can comprise validating the efficacy of a J peptide candidate such as any one of the 2O30candidates for a 30- J peptide. In an aspect, validating the efficacy of a disclosed J peptide candidate can comprise performing any one of the methods identified in the Examples (e.g., affinity chromatography, iodixanol density centrifugation, etc.).

[0322] In an aspect, a disclosed method can comprise validating the loading of a J peptide fused to a desired cargo into a disclosed AAV particle or AAV-like particle. In an aspect, validating theloading of a J peptide fused to a desired cargo can comprise performing any one of the methods identified in the Examples.2. Methods of Treating a Subject and / or Slowing Progression of a Disease or Disorder

[0323] Disclosed herein is a method of treating a subject, the method comprising: administering to a subject in need of a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV-like particle (or a composition thereof) carrying a disclosed cargo, wherein the delivery of the cargo to one or more cells or tissues in the subject provides a therapeutic effect.

[0324] Disclosed herein is a method of slowing progression of a disease or disorder in a subject, the method comprising: administering to a subject in need of a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV-like particle (or a composition thereof) carrying a disclosed cargo, wherein the delivery of the cargo to one or more cells or tissues in the subject provides a therapeutic effect.

[0325] Disclosed herein is a method, the method comprising: administering a disclosed AAV particle or a disclosed AAV-like particle to a subject in need thereof, wherein the AAV particle or AAV-like particle comprises a cargo that can provide a therapeutic effect to one or more cells, one or more tissues, and / or one or more organs in the subject.

[0326] Disclosed herein is a method of treating a subject, the method comprising: administering to a subject in need of a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV-like particle (or a composition thereof) carrying a disclosed cargo, wherein following the administering step, the cargo is delivered a target cell and / or target tissue in the subject, and wherein the delivery of the cargo to one or more cells or tissues in the subject provides a therapeutic effect.

[0327] Disclosed herein is a method of slowing progression of a disease or disorder in a subject, the method comprising: administering to a subject in need of a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV-like particle (or a composition thereof) carrying a disclosed cargo, wherein following the administering step, the cargo is delivered a target cell and / or target tissue in the subject, and wherein the delivery of the cargo to one or more cells or tissues in the subject provides a therapeutic effect.

[0328] Disclosed herein is a method, the method comprising: administering a disclosed AAV particle or a disclosed AAV-like particle to a subject in need thereof, wherein the AAV particle or AAV-like particle comprises a cargo that can provide a therapeutic effect to one or more cells, one or more tissues, and / or one or more organs in the subject, wherein following the administering step, the cargo is delivered a target cell and / or target tissue in the subject.

[0329] In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV-like particle can be delivered via intravenous (IV) administration and can comprise a rangeof about 1 x IO10vg / kg to about 2 x 1014vg / kg. In an aspect, for example, a disclosed vector can be administered at a dose of about 1 x 1011to about 8 x 1013vg / kg or about 1 x 1012to about 8 x 1013vg / kg. In an aspect, a disclosed AAV particle or a disclosed AAV-like particle can be administered at a dose of about 1 x 1013to about 6 x 1013vg / kg. In an aspect, a disclosed AAV particle or a disclosed AAV-like particle can be administered at a dose of at least about 1 x IO10, at least about 5 x IO10, at least about 1 x 1011, at least about 5 x 1011, at least about 1 x 1012, at least about 5 x 1012, at least about 1 x 1013, at least about 5 x 1013, or at least about 1 x 1014vg / kg. In an aspect, a disclosed AAV particle or a disclosed AAV-like particle can be administered at a dose of no more than about 1 x IO10, no more than about 5 x IO10, no more than about 1 x 1011, no more than about 5 x 1011, no more than about 1 x 1012, no more than about 5 x 1012, no more than about 1 x 1013, no more than about 5 x 1013, or no more than about 1 x 1014vg / kg. In an aspect, a disclosed AAV particle or a disclosed AAV-like particle can be administered at a dose of about 1 x 1012vg / kg. In an aspect, a disclosed AAV particle or a disclosed AAV-like particle can be administered at a dose of about 1 x 1011vg / kg. In an aspect, a disclosed AAV particle or a disclosed AAV-like particle can be administered in a single dose, or in multiple doses (such as 2, 3, 4, 5, 6, 7, 8, 9 or 10 doses) as needed for the desired therapeutic results. In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV-like particle can comprise a range determined by a skilled person.

[0330] In an aspect, administration of a disclosed AAV particle or a disclosed AAV-like particle carrying a cargo to a subject can comprise intravenous injection, intramuscular injection, intracoronary injection, intracerebroventricular injection, intracisterna magna injection, intracerebral injection, intrathecal injection, or any combination thereof.

[0331] In an aspect, a subject can be an adult, a child, or an infant. In an aspect, a subject can be treatment-naive (e.g., not having received therapeutic AAV particles or AAV-like particles).

[0332] In an aspect, a disclosed method can restore the balance of the subject’s metabolism. In an aspect, a disclosed method can restore one or more aspects of the subject’s metabolism. In an aspect, a disclosed method can correct or restore one or more aspects of the subject’s metabolism. In an aspect, a disclosed method can correct, restore, supplement, and / or replenish the enzymatic activity of one or more disclosed defective and / or deficient and / or missing enzymes. In an aspect, restoring the activity and / or expression and / or functionality of a disclosed defective and / or deficient and / or missing enzyme can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a controllevel (e.g., a level in a subject not having a defective and / or deficient and / or missing enzyme). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity and / or functionality is similar to that of a wild-type or control level.

[0333] In an aspect, a disclosed method can comprise restoring one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell types (such as, for example, liver cells and muscle cells); (ii) normalizing aspects of the autophagy pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or preserving organelle functionality and / or structural integrity; (v) preventing, slowing, and / or eliminating hypoglycemia, ketosis, and / or other liver abnormalities; (vi) correcting liver enzyme dysregulation; (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the multi -systemic manifestations of a disclosed disease or disorder; (viii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the disclosed disease or disorder, or (ix) any combination thereof. In an aspect, restoring one or more aspects of cellular homeostasis can comprise improving, enhancing, restoring, and / or preserving one or more aspects of cellular structural and / or functional integrity.

[0334] In an aspect of a disclosed method, techniques to monitor, measure, and / or assess the restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise qualitative (or subjective) means as well as quantitative (or objective) means. These means are known to the skilled person. For example, hypoglycemia can be sporadically and / or continuously measured and monitored. Methods and techniques for measuring and monitoring hypoglycemia are known to the skilled person and include, but not limited to, by continuous glucose monitoring (CGM) methods and capillary blood glucose sticks.

[0335] In an aspect, a disclosed method can comprise administering to the subject one or more additional therapeutic agents. In an aspect, a disclosed therapeutic agent can comprise enzyme replacement therapy, gene therapy, mRNA therapy, small molecule therapy, substrate reduction therapy, or any combination thereof.

[0336] In an aspect, a disclosed method can comprise treating the subject. In an aspect, treating the subject can comprise administering to the subject one or more agents that modulate the level of one or more differentially present cellular metabolites.

[0337] In an aspect, a disclosed method can comprise validating the efficacy of the administered AAV particle or AAV-like particle. In an aspect, validating the efficacy of the administered AAVparticle or AAV-like particle can comprise administering to the subject a disclosed AAV particle or AAV-like particle, measuring the activity or expression of one or more biomarkers related to cellular function; and comparing the resulting activity or expression level of the one or more biomarkers to a control level, wherein the administered AAV particle or AAV-like particle is effective when the activity or expression level of the one or more biomarkers following treatment is modulated compared to the control level. In an aspect, modulated can comprise increasing the activity or expression level of the one or more biomarkers. In an aspect, modulated can comprise increasing the activity or expression level of the one or more biomarkers. In an aspect, modulated can comprise decreasing the activity or expression level of the one or more biomarkers.

[0338] In an aspect, a disclosed method can comprise measuring one or more biomarkers prior to the administering of one or more disclosed polypeptides. In an aspect, a disclosed method can comprise measuring one or more biomarkers during the administering of one or more disclosed polypeptides. In an aspect, a disclosed method can comprise measuring one or more biomarkers after the administering of one or more disclosed polypeptides.

[0339] Thus, in an aspect, the increase or decrease in expression and / or activity level postadministration can be a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of reduction in between as compared to native or control levels (e.g., pre-administration level). In an aspect, the increase or decrease in expression and / or activity level post-administration can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% as compared to a native or control level (e.g., pre-administration level). In an aspect, the inhibition or reduction can be 0-25%, 25-50%, 50-75%, or 75-100% as compared to native or control levels (e.g., pre- administration level). In an aspect, a native or control level can be a pre-disease or pre-disorder level or pre-treatment level).

[0340] In an aspect, measuring the expression of one or more disclosed biomarkers can comprise measuring the protein concentration of the disclosed cargo or measuring the mRNA level of disclosed cargo. For example, in an aspect, measuring the protein concentration of disclosed cargo can comprise a protein chip analysis, an immunoassay, a ligand binding assay, a MALDI-TOF (Matrix Assisted Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, a SELDLTOF (Sulface Enhanced Laser Desorption / Ionization Time of Flight Mass Spectrometry) analysis, a radioimmunoassay, a radioimmunodiffusion assay, an octeroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, a complement fixation assay, 2D by electrophoretic analysis, liquid chromatography-Mass Spectrometry (LC-MS), liquid chromatography -Mass Spectrometry / Mass Spectrometry (LC-MS / MS), Western blotting, ELISA (enzyme linked immunosorbent assay), or any combination thereof. Similarly, in an aspect, measuring the mRNA level of a disclosed cargo can comprise a reverse transcription polymerasereaction (RT-PCR), a competitive reverse transcription polymerase reaction (Competitive RT- PCR), a real-time reverse transcription polymerization, an enzyme reaction (Real-time RT-PCR), an RNase protection assay (RPA), Northern blotting, a DNA chip, or any combination thereof.

[0341] In an aspect, a disclosed method can further comprise monitoring the subject for adverse effects. In an aspect, in the absence of adverse effects, the method can further comprise continuing to treat the subject. In an aspect, in the presence of adverse effects, the method can further comprise modifying the treating step. Methods of monitoring a subject’s well-being can include both subjective and objective criteria. Such methods are known to the skilled person. In an aspect, a disclosed method can further comprise repeating a monitoring step.

[0342] In an aspect, a disclosed method of treating and / or preventing disease progression in a subject can further comprise administering one or more immune modulators. In an aspect, a disclosed immune modulator can be methotrexate, rituximab, intravenous gamma globulin, or bortezomib, or a combination thereof. In an aspect, a disclosed immune modulator can be bortezomib or SVP-Rapamycin. In an aspect, a disclosed immune modulator can be Tacrolimus. In an aspect, a disclosed immune modulator such as methotrexate can be administered at a transient low to high dose. In an aspect, a disclosed immune modulator can be administered at a dose of about 0.1 mg / kg body weight to about 0.6 mg / kg body weight. In an aspect, a disclosed immune modulator can be administered at a dose of about 0.4 mg / kg body weight. In an aspect, a disclosed immune modulator can be administered at about a daily dose of 0.4 mg / kg body weight for 3 to 5 or greater cycles, with up to three days per cycle. In an aspect, a disclosed immune modulator can be administered at about a daily dose of 0.4 mg / kg body weight for a minimum of 3 cycles, with three days per cycle. In an aspect, a person skilled in the art can determine the appropriate number of cycles. In an aspect, a disclosed immune modulator can be administered as many times as necessary to achieve a desired clinical effect.

[0343] In an aspect, a disclosed immune modulator can be administered orally about one hour before a disclosed therapeutic agent. In an aspect, a disclosed immune modulator can be administered subcutaneously about 15 minutes before a disclosed therapeutic agent. In an aspect, a disclosed immune modulator can be administered concurrently with a disclosed therapeutic agent. In an aspect, a disclosed immune modulator can be administered orally about one hour or a few days before a disclosed polypeptide, a disclosed isolated nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, or a combination thereof. In an aspect, a disclosed immune modulator can be administered subcutaneously about 15 minutes before or a few days before a disclosed polypeptide, a disclosed isolated nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, or a combination thereof. In an aspect, a disclosed immune modulator can be administered concurrently with a disclosed polypeptide, a disclosed isolatednucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, or a combination thereof.

[0344] In an aspect, a disclosed method of treating and / or preventing disease progression in a subject can further comprise administering one or more proteasome inhibitors (e.g., bortezomib, carfilzomib, marizomib, ixazomib, and oprozomib). In an aspect, a proteasome inhibitor can be an agent that acts on plasma cells (e.g., daratumumab). In an aspect, an agent that acts on a plasma cell can be melphalan hydrochloride, melphalan, pamidronate disodium, carmustine, carfilzomib, carmustine, cyclophosphamide, daratumumab, doxorubicin hydrochloride liposome, doxorubicin hydrochloride liposome, elotuzumab, melphalan hydrochloride, panobinostat, ixazomib citrate, carfilzomib, lenalidomide, melphalan, melphalan hydrochloride, plerixafor, ixazomib citrate, pamidronate disodium, panobinostat, plerixafor, pomalidomide, pomalidomide, lenalidomide, selinexor, thalidomide, thalidomide, bortezomib, selinexor, zoledronic acid, or zoledronic acid.

[0345] In an aspect, a disclosed method can further comprise administering one or more proteasome inhibitors or agents that act on plasma cells prior to administering a disclosed polypeptide, a disclosed isolated nucleic acid molecule, a disclosed vector, or a disclosed pharmaceutical formulation. In an aspect, a disclosed method can comprise administering one or more proteasome inhibitors or one or more agents that act on plasma cells concurrently with administering a disclosed AAV particle, a disclosed AAV-like particle, a disclosed polypeptide, a disclosed nucleic acid molecule, a disclosed vector, or a disclosed pharmaceutical formulation. In an aspect, a disclosed method can comprise administering one or more proteasome inhibitors or one or more agents that act on plasma cells subsequent to administering a disclosed polypeptide, a disclosed isolated nucleic acid molecule, a disclosed vector, or a disclosed pharmaceutical formulation. In an aspect, a disclosed method can further comprise administering one or more proteasome inhibitors more than 1 time. In an aspect, a disclosed method can comprise administering one or more proteasome inhibitors repeatedly over time.

[0346] In an aspect, a disclosed method of treating and / or preventing disease progression in a subject can further comprise administering one or more immunosuppressive agents. In an aspect, an immunosuppressive agent can be, but is not limited to, azathioprine, methotrexate, sirolimus, anti -thymocyte globulin (ATG), cyclosporine (CSP), mycophenolate mofetil (MMF), steroids, or a combination thereof. In an aspect, a disclosed method can comprise administering one or more immunosuppressive agents more than 1 time. In an aspect, a disclosed method can comprise administering one or more one or more immunosuppressive agents repeatedly over time. In an aspect, a disclosed method can comprise administering a compound that targets or alters antigen presentation or humoral or cell mediated or innate immune responses.

[0347] In an aspect, a disclosed method can further comprise administering a compound that exerts a therapeutic effect against B cells and / or a compound that targets or alters antigen presentation or humoral or cell mediated immune response. In an aspect, a disclosed compound can be rituximab, methotrexate, intravenous gamma globulin, anti CD4 antibody, anti CD2, an anti-FcRN antibody, a BTK inhibitor, an anti -IGF 1R antibody, a CD 19 antibody (e.g., inebilizumab), an anti-IL6 antibody (e.g., tocilizumab), an antibody to CD40, an IL2 mutein, or a combination thereof. Also disclosed herein are Treg infusions that can be administered as a way to help with immune tolerance (e.g., antigen specific Treg cells to AAV).

[0348] In an aspect, a disclosed method of treating and / or preventing disease progression in a subject can further comprise repeating a disclosed administering step such as, for example, repeating the administering of a disclosed AAV particle, a disclosed AAV-like particle, a disclosed plasmid, a disclosed polypeptide, a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, a disclosed therapeutic agent, a disclosed immune modulator, a disclosed proteasome inhibitor, a disclosed immunosuppressive agent, a disclosed compound that exerts a therapeutic effect against B cells and / or a disclosed compound that targets or alters antigen presentation or humoral or cell mediated immune response.

[0349] In an aspect, a disclosed method of treating and / or preventing disease progression in a subject can further comprise modifying one or more of the disclosed steps. For example, modifying one or more of steps of a disclosed method can comprise modifying or changing one or more features or aspects of one or more steps of a disclosed method. For example, in an aspect, a method can be altered by changing the amount of a disclosed AAV particle, a disclosed AAV- like particle, a disclosed polypeptide or a pharmaceutical formulation thereof administered to a subject, or by changing the frequency of administration of a disclosed AAV particle, a disclosed AAV-like particle, a disclosed polypeptide or a pharmaceutical formulation thereof to a subject, or by changing the duration of time a disclosed AAV particle, a disclosed AAV-like particle, a disclosed polypeptide or a pharmaceutical formulation thereof is administered to a subject. In an aspect, one or more disclosed AAV particles or AAV-like particles can be administered concurrently or sequentially.

[0350] In an aspect, a disclosed method can further comprise diagnosing a subject with a genetic defect using one or more known methods to the skilled person, such as, for example, genotyping. In an aspect...

Claims

VIII. CLAIMSWhat is claimed is:

1. A peptide, comprising: the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29.

2. The peptide of Claim 1, wherein the peptide binds to the luminal surface of the capsid of an adeno-associated virus (AAV) or an AAV-like particle.

3. The peptide of Claim 1, wherein the peptide is fused to a cargo via a linker.

4. The peptide of Claim 3, wherein the linker comprises the sequence set forth in SEQ ID NO: 185 or SEQ ID NO: 186.

5. The peptide of Claim 3, wherein the cargo comprises a protein or enzyme.

6. The peptide of Claim 4, wherein the protein or enzyme comprises a protein or an enzyme that is missing, deficient, and / or mutated in one or more cells in a subject.

7. The peptide of Claim 3, wherein the cargo comprises an RNA binder, a DNA binder, an epigenetic modulator for activation or repression, an inhibitor of a cellular degradation process, an antibody or an antibody fragment, or a CRISPR / Cas protein.

8. A nucleic acid molecule, comprising: a nucleic acid sequence encoding the peptide of any one of Claims 1 - 2.

9. The nucleic acid molecule of Claim 8, wherein the nucleic acid sequence encoding the peptide comprises the sequence set forth in any one of SEQ ID NO:49 - SEQ ID NO:78.

10. The nucleic acid molecule of any one of Claims 8 - 9, further comprising a linker sequence.

11. The nucleic acid molecule of Claim 10, wherein the linker sequence comprises the sequence set forth in any one of SEQ ID NO:193 - SEQ ID NO:213.

12. The nucleic acid molecule of any one of Claims 8 - 11, further comprising a nucleic acid sequence for a cargo.

13. The nucleic acid molecule of Claim 12, wherein the cargo comprises a protein or enzyme.

14. The nucleic acid molecule of Claim 13, wherein the protein or enzyme comprises a protein or an enzyme that is missing, deficient, and / or mutated in one or more cells in a subject.

15. The nucleic acid molecule of Claim 12, wherein the cargo comprises an RNA binder, a DNA binder, an epigenetic modulator for activation or repression, an inhibitor of a cellular degradation process, an antibody or an antibody fragment, or a CRISPR / Cas protein.

16. An AAV particle, comprising: the peptide of any one of Claims 2 - 7 or the nucleic acid molecule of any one of Claims 8 - 15.

17. An AAV-like particle, comprising: the peptide of any one of Claims 2 - 7 or the nucleic acid molecule of any one of Claims 8 - 15.

18. A pharmaceutical formulation, comprising: (i) the AAV particle of Claim 16 and one or more pharmaceutically acceptable carriers; or (ii) the AAV-like particle of Claim 17 and one or more pharmaceutically acceptable carriers.

19. A method of treating a subject, the method comprising: administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical formulation of Claim 18, wherein following the administering step, the cargo is delivered to one or more target cells and / or target tissues in the subject.

20. The method of Claim 19, wherein the delivery of the cargo to one or more target cells or target tissues in the subject provides a therapeutic effect21. The method of Claim 19, wherein the subject has been diagnosed with or is suspected of having a genetic disease or disorder.

22. The method of Claim 19 or Claim 20, wherein following the administering step, one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation are restored in one or more cells, tissues, or organs in the subject.

23. The method of any one of Claims 19 - 22, further repeating the administering step.

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