Cas6 polypeptides and methods of use
AssigneeUniversity of Georgia Research Foundation, Inc.
InventorsRebecca M. Terns, Michael P. Terns, Jason Carte
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Abstract
Provided herein are methods for cleaving a target RNA polynucleotide. The target RNA polynucleotide includes a Cas6 recognition domain and a cleavage site, and may be based on a repeat from a CRISPR locus. The methods may be practiced in vivo or in vitro. Also provided are polypeptides that have Cas6 endoribonuclease activity in the presence of a target RNA polynucleotide, and methods for using the polypeptides.
Claims20
-6. (canceled)
A genetically modified microbe comprising an exogenous polynucleotide, wherein the exogenous polynucleotide comprises a nucleotide sequence encoding a polypeptide having Cas6 endoribonuclease activity, wherein the amino acid sequence of the polypeptide and the amino acid sequence of SEQ ID NO:2 have at least 80% identity.
An enriched polypeptide having Cas6 endoribonuclease activity, wherein the polypeptide comprises an amino acid sequence, wherein the amino acid sequence and the amino acid sequence of SEQ ID NO:2 have at least 80% identity.
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12-19. (canceled)
A method for cleaving a nucleotide sequence comprising:
- incubating a target RNA polynucleotide with a polypeptide under conditions suitable for cleavage of the target RNA polynucleotide,
- wherein the target RNA polynucleotide comprises a Cas6 recognition domain present in a prokaryotic genome;
- wherein the polypeptide comprises an amino acid sequence of a Cas6 polypeptide from the prokaryotic genome and has Cas6 endoribonuclease activity;
- and
- wherein the polypeptide cleaves the target RNA polynucleotide, the cleavage site located 5 to 20 nucleotides downstream of the Cas6 endoribonuclease domain.
23-28. (canceled)
Dependencies are inferred from the claim text.