Virginijus {hacek over (S)}ik{hacek over (s)}nys,Giedrius Gasiunas,Tautvydas Karvelis
申请号：
US14385241
公开号：
US09637739B2
申请日:
2013.03.15
申请国别(地区):
US
年份:
2017
代理人:
摘要：
Isolation or in vitro assembly of the Cas9-crRNA complex of the Streptococcus thermophilus CRISPR3/Cas system and use for cleavage of DNA bearing a nucleotide sequence complementary to the crRNA and a proto-spacer adjacent motif. Methods for site-specific modification of a target DNA molecule using 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； for conversion of Cas9 polypeptide into a nickase cleaving one strand of double-stranded DNA by inactivating one of the active sites (RuvC or HNH) in the polypeptide by at least one point mutation； for assembly of active polypeptide-polyribonucleotides complex in vivo or in vitro； and for re-programming a Cas9-crRNA complex specificity in vitro or using a cassette containing a single repeat-spacer-repeat unit.
