Rna-guided nucleases and dna binding proteins
AssigneeInari Agriculture Technology, Inc.
InventorsNeena Kenton Pyzocha, Adam Patrick Joyce, Karl Kremling
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Abstract
Compositions and methods related to Cas proteins, nucleic acids encoding the Cas proteins, and modified host cells comprising the Cas proteins and/or encoding nucleic acids are disclosed. Cas proteins are useful in a variety of applications. Cas proteins bind guide RNAs that in turn provide functional specificity to the Cas proteins, nucleic acids encoding the Cas guide RNAs, and modified host cells comprising the Cas guide RNAs and/or encoding nucleic acids. The Cas polypeptides and corresponding guide RNAs can be used in a variety of applications.
Claims128
A composition comprising:
- a) a CasP6 polypeptide, or a nucleic acid molecule encoding the CasP6 polypeptide;
- and
- b) a CasP6 guide RNA, or one or more DNA molecules encoding the CasP6 guide RNA.
A CasP6 single guide RNA molecule, comprising:
- a) a guide sequence that hybridizes to a target nucleic acid, and a duplex-forming segment;
- and
- b) an activator sequence that can bind a CasP6 polypeptide.
A CasP6 fusion polypeptide comprising:
- a CasP6 polypeptide fused to a heterologous polypeptide.
One or more nucleic molecules encoding:
- (a) a CasP6 guide RNA comprising an activator RNA and a guide RNA;
- and
- (b) a CasP6 polypeptide.
A eukaryotic cell comprising one or more of:
- a) a CasP6 polypeptide, or a nucleic acid molecule encoding the CasP6 polypeptide,
- b) a CasP6 guide RNA, or a nucleic acid molecule encoding the CasP6 guide RNA.
A cell comprising a CasP6 fusion polypeptide, or a nucleic acid molecule encoding the CasP6 fusion polypeptide.
A method of modifying a target nucleic acid, the method comprising contacting the target nucleic acid with:
- a) a CasP6 polypeptide;
- and
- b) a CasP6 guide RNA comprising a guide sequence that hybridizes to a target sequence of the target nucleic acid,
- wherein said contacting results in modification of the target nucleic acid by the CasP6 polypeptide.
A method of modulating transcription from a target DNA, modifying a target nucleic acid, or modifying a protein associated with a target nucleic acid, the method comprising contacting the target nucleic acid with:
- a) a CasP6 fusion polypeptide comprising a CasP6 polypeptide fused to a heterologous polypeptide;
- and
- b) a CasP6 guide RNA comprising a guide sequence that hybridizes to a target sequence of the target nucleic acid.
A transgenic, multicellular, non-human organism whose genome comprises a transgene comprising a nucleotide sequence encoding one or more of:
- a) a CasP6 polypeptide, and/or
- b) a CasP6 guide RNA.
A system comprising:
- a) a CasP6 polypeptide and a CasP6 single guide RNA;
- b) a CasP6 polypeptide, a CasP6 guide RNA, and a DNA donor template;
- c) a CasP6 fusion polypeptide and a CasP6 guide RNA;
- d) a CasP6 fusion polypeptide, a CasP6 guide RNA, and a DNA donor template;
- e) an mRNA encoding a CasP6 polypeptide, and a CasP6 single guide RNA;
- f) an mRNA encoding a CasP6 polypeptide;
- a CasP6 guide RNA, and a DNA donor template;
- g) an mRNA encoding a CasP6 fusion polypeptide, and a CasP6 guide RNA;
- h) an mRNA encoding a CasP6 fusion polypeptide, a CasP6 guide RNA, and a DNA donor template;
- i) one or more recombinant expression vectors comprising:
- i) a nucleotide sequence encoding a CasP6 polypeptide;
- and ii) a nucleotide sequence encoding a CasP6 guide RNA;
- j) one or more recombinant expression vectors comprising:
- i) a nucleotide sequence encoding a CasP6 polypeptide;
- ii) a nucleotide sequence encoding a CasP6 guide RNA;
- and iii) a DNA donor template;
- k) one or more recombinant expression vectors comprising:
- i) a nucleotide sequence encoding a CasP6 fusion polypeptide;
- and ii) a nucleotide sequence encoding a CasP6 guide RNA;
- or
- l) one or more recombinant expression vectors comprising:
- i) a nucleotide sequence encoding a CasP6 fusion polypeptide;
- ii) a nucleotide sequence encoding a CasP6 guide RNA;
- and a DNA donor template.
Dependencies are inferred from the claim text.