METHOD FOR INDUCING HOMOLOGOUS RECOMBINATION USING CAS9 NUCLEASE

摘要

The present invention provides: (1) a method for inducing homologous recombination between the genomic DNA of a cell and a donor DNA containing an insertion sequence into the genomic DNA , said method comprising a step for contacting the genomic DNA, the donor DNA and Cas9 nuclease in the absence of guide RNA; and (2) a method for evaluating the possibility of off-target mutagenesis by a nucleic acid for modifying the genomic DNA of a cell, said method comprising a step for calculating the degree of homology between at least a partial sequence of the nucleic acid and at least a partial sequence of the the genomic DNA of the cell, wherein the nucleic acid for modifying the genomic DNA contains one or more nucleotide sequences selected from the group consisting of a nucleotide sequence that encodes a nucleic acid sequence-recognizing module specifically binding to a target nucleotide sequence in the genomic DNA of the cell, a nucleotide sequence that encodes a nucleic acid modifying enzyme, and an insertion sequence into the genomic DNA of the cell.