Redirection of tropism of AAV capsids
AssigneeVoyager Therapeutics, Inc.
InventorsMathieu E. Nonnenmacher, Jinzhao Hou, Wei Wang
Life of this patent
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Abstract
The disclosure relates to compositions, methods, and processes for the preparation, use, and/or formulation of adeno-associated virus capsid proteins, wherein the capsid proteins comprise targeting peptide inserts for enhanced tropism to a target tissue.
Claims29
A method for generating an adeno-associated virus (AAV) vector library encoding variant AAV capsid polypeptides, the method comprising:
- (a) providing first nucleic acids comprising a P40 promoter and a cell-type specific promoter, wherein the cell-type specific promoter drives capsid mRNA expression in the absence of helper virus co-infection;
- and second nucleic acids encoding the variant AAV capsid polypeptides having a region of randomized sequence of 2, 3, 4, 5, 6, 7, 8, or 9 consecutive amino acids;
- and
- (b) cloning the first nucleic acids and the second nucleic acids under conditions suitable to generate the AAV vector library.
A method for generating an adeno-associated virus (AAV) vector library encoding variant AAV capsid polypeptides, the method comprising:
- (a) providing first nucleic acids comprising a first promoter and a second promoter, wherein the second promoter is a cell-type specific promoter which drives capsid mRNA expression in the absence of helper virus co-infection;
- and second nucleic acids encoding the variant AAV capsid polypeptides having a region of randomized sequence of 2, 3, 4, 5, 6, 7, 8, or 9 consecutive amino acids;
- and
- (b) cloning the first nucleic acids and the second nucleic acids under conditions suitable to generate the AAV vector library;
- wherein the cell-type specific promoter is a neuron specific promoter, an astrocyte-specific promoter, a blood cell specific promoter, an eye specific promoter, a heart specific promoter, a muscle specific promoter, a kidney specific promoter, a lung specific promoter, a pancreas specific promoter, or a vasculature specific promoter.
A method for generating a plurality of variant adeno-associated virus (AAV) capsid polypeptides, the method comprising:
- (a) providing a plurality of AAV particles comprising an AAV vector library, wherein the AAV vector library comprises nucleic acids comprising a P40 promoter and a cell-type specific promoter which drives capsid mRNA expression in the absence of helper virus co-infection;
- and
- wherein the nucleic acids encode a plurality of the variant AAV capsid polypeptides having a region of randomized sequence of 2, 3, 4, 5, 6, 7, 8, or 9 consecutive amino acids;
- (b) administering the AAV particles to an animal, wherein the particles are administered intravenously;
- (c) recovery of RNA and/or antisense RNA encoding the variant AAV capsid polypeptides from a target cell or tissue,
- (d) determination of the sequence of the variant AAV capsid polypeptides;
- and
- (e) measuring the amount of DNA encoding the variant AAV capsid polypeptides or the amount of RNA encoding the variant AAV capsid polypeptides, in a target cell or tissue;
- and
- wherein the method occurs in the absence of a helper virus;
- thereby generating the plurality of variant AAV capsid polypeptides.
Dependencies are inferred from the claim text.