Targeted gene modification by parvoviral vectors

摘要

This invention provides methods for obtaining targeted gene modification in vertebrate cells using parvoviral vectors, including adeno-associated virus (AAV). The parvoviral vectors used in the methods of the invention are capable of targeting a specific genetic modification to a preselected target locus in a cellular genome by homologous pairing.

主权项

1. A recombinant, replication-defective adeno-associated virus (AAV) viral particle for introducing a genetic modification of a preselected target gene in the genome of a vertebrate cell by homologous recombination, the recombinant, replication-defective AAV viral particle comprising:
a targeting construct comprising a genomic DNA sequence which is substantially identical to the preselected target gene sequence except for the modification to be introduced; wherein the genomic DNA sequence consists of less than an entire copy of the preselected target gene; wherein the modification to be introduced is flanked by two regions of nucleotide sequence which are substantially identical to the preselected target gene sequence, each region being at least 100 nucleotides in length; wherein the targeting construct comprises two AAV inverted terminal repeats (ITRs), one at each end of the targeting construct; wherein the modification comprises one or more insertions or substitutions or a combination thereof; and wherein the viral particle introduces the modification into the preselected target gene by homologous recombination at a frequency of at least 0.01%.