1 . A in-vitro method for the site-specific modification of a target DNA molecule, the method comprising
contacting, under suitable conditions,
a target DNA molecule; and
an RNA-guided DNA endonuclease comprising at least one RNA sequence and at least one of an RuvC active site motif and an HNH active site motif;
wherein
the RNA-guided DNA endonuclease is a Cas9-crRNA complex
said Cas9-crRNA complex comprising a Cas9, crRNA and tracrRNA
to result in the target DNA molecule modified in a region that is determined by the complimentary binding of the crRNA sequence to the target DNA molecule,
the method further comprising assembling the popeptide-polyribonucleotides complex in-vitro by incubating said components of the complex under conditions suitable for complex assembly,
wherein the target DNA is double stranded or single stranded and wherein a double stranded target DNA contains a proto-spacer adjacent motif.
・日米欧共にin vitro assemblyが必須なので、特許としては…
