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US 11,692,184 B2Granted
Thermostable RNA-guided endonucleases and methods of use thereof
AssigneeThe Regents of the University of California
InventorsNikos C. Kyrpides, Jennifer A. Doudna, Lucas Benjamin Harrington, David Paez-Espino
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Abstract
The present disclosure provides RNA-guided endonucleases, nucleic acids encoding same, and compositions comprising same. The present disclosure provides ribonucleoprotein complexes comprising: an RNA-guided endonuclease of the present disclosure; and a guide RNA. The present disclosure provides methods of modifying a target nucleic acid, using an RNA-guided endonuclease of the present disclosure and a guide RNA.
Claims16
An RNA-guided endonuclease that comprises:
- a) an amino acid sequence having at least 95% amino acid sequence identity to amino acids 1-994 of the amino acid sequence set forth in SEQ ID NO:
- 1;
- and
- b) a heterologous protospacer adjacent motif (PAM) interacting (PI) domain comprising an amino acid sequence having at least 95% amino acid sequence identity to the following amino acid sequence:
- IIKTAGGEEIKIKDLFAYYKTIHSGTAGLELVSHDCSFSLSGVGSRTLKRFEKYQVDVLGNIYKVRGEKRVGLASSAHSKTGETIRPLQSTRD (SEQ ID NO:
- 4), wherein the PI domain is heterologous to the amino acid sequence of part a).
3 more elements
A fusion polypeptide comprising:
- a) an RNA-guided endonuclease that comprises:
- i) an amino acid sequence having at least 95% amino acid sequence identity to amino acids 1-994 of the amino acid sequence set forth in SEQ ID NO:
- 1;
- and
- ii) a heterologous protospacer adjacent motif (PAM) interacting (PI) domain comprising an amino acid sequence having at least 95% amino acid sequence identity to the following amino acid sequence:
- IIKTAGGEEIKIKDLFAYYKTIHSGTAGLELVSHDCSFSLSGVGSRTLKRFEKYQVDVLGNIYKVRGEKRVGLASSAHSKTGETIRPLQSTRD (SEQ ID NO:
- 4), wherein the PI domain is heterologous to the amino acid sequence of part i);
- and
- b) a heterologous fusion partner polypeptide.
6 more elements
Dependencies are inferred from the claim text.