# US 11,028,381 B2 · CRISPR-associated (Cas) protein > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,028,381 B2 - Document ID: US11028381B2 - Status: Granted - Kind: B2 - Application number: 16/693,182 - Filed: 2019-11-22 - Published: 2020-03-12 - Granted: 2021-06-08 - Earliest claimed: 2017-03-28 - Assignees: Locanabio, Inc. - Applicants: Locanabio, Inc. - Inventors: Matthew Merrill Carter; Paul Daniel Donohoue - Web page: https://patentgrep.com/patent/US11028381B2 ## CPC - C12N 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} - C12N 9/222: {Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes} - C12N 15/11: DNA or RNA fragments; Modified forms thereof (DNA or RNA not used in recombinant technology, C07H21/00); {Non-coding nucleic acids having a biological activity} - C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids} - C12N 15/1136: {against growth factors, growth regulators, cytokines, lymphokines or hormones} - C12N 15/85: for animal cells - C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR] - C12N 2800/22: Vectors comprising a coding region that has been codon optimised for expression in a respective host - Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract A new CRISPR-associated (Cas) protein, termed “CasM,” is described, as well as polynucleotides encoding the same and methods of using CasM for site-specific genome engineering. CasM proteins are capable of targeting and cleaving single-stranded RNA. ## Claims ### Claim 1 A method of directing a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) nucleoprotein complex to a selected nucleic acid target sequence, the method comprising: contacting the selected nucleic acid target sequence with one or more nucleoprotein complexes comprising a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) protein, wherein the Cas protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:44, an amino acid sequence having at least 98 percent sequence identity to SEQ ID NO:37, an amino acid sequence having at least 98 percent sequence identity to SEQ ID NO:39, and an amino acid sequence having at least 98 percent sequence identity to SEQ ID NO:44; and a cognate nucleic acid guide comprising a repeat sequence and a spacer sequence, wherein the repeat sequence and the spacer sequence do not naturally occur together, wherein the Cas protein and the cognate nucleic acid guide form the nucleoprotein complex, and wherein the nucleoprotein complex binds to the selected nucleic acid target sequence. ### Claim 2 (depends on 1) The method of claim 1, wherein the nucleic acid target sequence comprises RNA. ### Claim 3 (depends on 1) The method of claim 1, wherein the method is performed in a cell. ### Claim 4 (depends on 3) The method of claim 3, wherein the cell is a eukaryotic cell. ### Claim 5 (depends on 3) The method of claim 3, wherein the cell constitutively expresses the Cas protein. ### Claim 6 (depends on 1) The method of claim 1, wherein the Cas protein is fused to a protein. ### Claim 7 (depends on 6) The method of claim 6, wherein the protein which is fused to the Cas protein comprises an enzyme. ### Claim 8 (depends on 1) The method of claim 1, wherein the cognate nucleic acid guide comprises RNA. ### Claim 9 (depends on 1) The method of claim 1, wherein the cognate nucleic acid guide comprises DNA. ### Claim 10 (depends on 1) The method of claim 1, wherein the Cas protein comprises an amino acid sequence having at least 98 percent sequence identity to SEQ ID NO:44. ### Claim 11 (depends on 1) The method of claim 1, wherein the nucleoprotein complex is capable of site-directed binding to the nucleic acid target sequence. ### Claim 12 (depends on 11) The method of claim 11, wherein the Cas protein is catalytically active, resulting in the binding of the nucleoprotein complex to the nucleic acid target sequence and cleaving of the nucleic acid target sequence. ### Claim 13 (depends on 12) The method of claim 12, whereby the cleaving results in one or more single-strand or double-strand breaks in the nucleic acid target sequence. ### Claim 14 (depends on 12) The method of claim 12, wherein the nucleic acid target sequence comprises RNA. ### Claim 15 (depends on 12) The method of claim 12, wherein the method is performed in a cell. ### Claim 16 (depends on 15) The method of claim 15, wherein the cell constitutively expresses the Cas protein. ### Claim 17 (depends on 15) The method of claim 15, wherein the cell comprises a eukaryotic cell. ### Claim 18 (depends on 17) The method of claim 17, wherein the eukaryotic cell comprises a mammalian cell. ### Claim 19 (depends on 18) The method of claim 18, wherein the mammalian cell comprises a human cell. ### Claim 20 (depends on 19) The method of claim 19, wherein the human cell comprises a stem cell. ### Claim 21 A method of directing a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) nucleoprotein complex to a selected nucleic acid target sequence, the method comprising: contacting the selected nucleic acid target sequence with one or more nucleoprotein complexes comprising a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) protein, wherein the Cas protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:37, SEQ ID NO:39, and SEQ ID NO:44; and a cognate nucleic acid guide comprising a repeat sequence and a spacer sequence, wherein the repeat sequence and the spacer sequence do not naturally occur together, wherein the Cas protein and the cognate nucleic acid guide form the nucleoprotein complex, and wherein the nucleoprotein complex binds to the selected nucleic acid target sequence. ### Claim 22 (depends on 21) The method of claim 21, wherein the Cas protein comprises the amino acid sequence of SEQ ID NO: 37. ### Claim 23 (depends on 21) The method of claim 21, wherein the Cas protein comprises the amino acid sequence of SEQ ID NO: 39. ### Claim 24 (depends on 21) The method of claim 21, wherein the Cas protein comprises the amino acid sequence of SEQ ID NO: 44. ### Claim 25 (depends on 1) The method of claim 1, wherein the Cas protein is catalytically inactive.