Isolation of novel AAV's and uses thereof
AssigneeUniversity of Massachusetts
InventorsGuangping Gao, Phillip D. Zamore
Life of this patent
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Abstract
The invention in some aspects relates to isolated nucleic acids, compositions, and kits useful for identifying adeno-associated viruses in cells. In some aspects, the invention provides kits and methods for producing somatic transgenic animal models using recombinant AAV (rAAV) to an animal having at least one transgene that expresses a small interfering nucleic acid or at least one binding site for a miRNA.
Claims16
A method for delivering a transgene to a target tissue in a subject, the method comprising administering to the subject a recombinant adeno-associated virus (rAAV) that transduces cells of the target tissue and a transgene, wherein the transgene is engineered to express an RNA transcript that comprises at least one binding site for a miRNA that is expressed in cells of an off-target tissue of the subject, wherein the at least one miRNA binding site is positioned in the 5′UTR or 3′UTR of the RNA transcript, wherein the transgene comprises a tissue-specific promoter selected from a thyroxin binding globulin (TBG) promoter, insulin promoter, glucagon promoter, somatostatin promoter, pancreatic polypeptide (PPY) promoter, synapsin-1 (Syn) promoter, creatine kinase (MCK) promoter, mammalian desmin (DES) promoter, α-myosin heavy chain (α-MHC) promoter, and a cardiac Troponin T (cTnT) promoter, and wherein the rAAV comprises a capsid protein from an AAV serotype selected from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV10, AAV11, or AAV12.
A method for inhibiting transgene expression in an off-target tissue, the method comprising administering to a subject a recombinant adeno-associated virus (rAAV) comprising a transgene that is engineered to express an RNA transcript that comprises at least one binding site for a miRNA that is expressed in cells of an off-target tissue of the subject, wherein the at least one miRNA binding site is positioned in the 5′UTR or 3′UTR of the RNA transcript, wherein the transgene comprises a tissue-specific promoter selected from a thyroxin binding globulin (TBG) promoter, insulin promoter, glucagon promoter, somatostatin promoter, pancreatic polypeptide (PPY) promoter, synapsin-1 (Syn) promoter, creatine kinase (MCK) promoter, mammalian desmin (DES) promoter, α-myosin heavy chain (α-MHC) promoter, and a cardiac Troponin T (cTnT) promoter, wherein the rAAV comprises a capsid protein from an AAV serotype selected from AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV10, AAV11, or AAV12, and wherein, when the miRNA and the RNA transcript are present together in the off-target tissue, the miRNA hybridizes to the at least one binding site of the RNA transcript and inhibits activity of the RNA transcript.
Dependencies are inferred from the claim text.