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US 2018/0002682 A1Published application
Heterodimeric cas9 and methods of use thereof
AssigneeThe Regents of the University of California
InventorsSamuel H. Sternberg, Jennifer A. Doudna, Addison V. Wright
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Abstract
The present disclosure provides a Cas9 heterodimer, as well as nucleic acids encoding the Cas9 heterodimer, and host cells comprising the nucleic acids. The present disclosure provides a system that includes a Cas9 heterodimer of the present disclosure and at least one of: a Cas9 guide RNA, and a dimerizing agent. A Cas9 heterodimer of the present disclosure is useful in a wide variety of applications, which are also provided.
Claims62
A Cas9 heterodimer comprising:
- A) a first fusion polypeptide comprising:
- a) a first polypeptide comprising:
- i) a RuvCI polypeptide;
- ii) a RuvCII polypeptide;
- iii) an HNH polypeptide;
- iv) a RuvCIII polypeptide;
- and
- v) a PAM-interacting polypeptide;
- and
- b) a first fusion partner, where the first fusion partner is a first member of a dimerization pair;
- and
- B) a second fusion polypeptide comprising:
- a) an alpha-helical recognition region;
- and
- b) a second fusion partner, where the second fusion partner is a second member of a dimerization pair.
12 more elements
A Cas9 heterodimer comprising:
- A) a first fusion polypeptide comprising:
- a) a first, circular permuted, polypeptide that comprises:
- i) a RuvCI polypeptide;
- ii) a RuvCII polypeptide;
- iii) an HNH polypeptide;
- iv) a RuvCIII polypeptide;
- and
- v) a PAM-interacting polypeptide;
- and
- b) a first fusion partner, where the first fusion partner is a first member of a dimerization pair;
- and
- B) a second fusion polypeptide comprising:
- a) a second polypeptide that comprises an alpha-helical recognition region;
- and
- b) a second fusion partner, where the second fusion partner is a second member of the dimerization pair.
12 more elements
Dependencies are inferred from the claim text.