Granted Patent B2

US 12,195,749 · App. 17/692,069 · Granted Jan 14, 2025

Inventors: Andrew Garst (Pleasanton, CA); Ryan T. Gill (Pleasanton, CA); Tanya Elizabeth Warnecke Lipscomb (Pleasanton, CA)

Assignee: Inscripta, Inc.

C12N15/902C12N9/22C12N15/11C12N15/111C12N15/70C12N15/81C12N15/85C12N15/905C12N15/907C12N15/8509C12N2310/20C12N2800/22

Abstract

Claims (25)

1. A nucleic acid-guided nuclease system comprising:

(a) a nucleic acid-guided nuclease encoded by a nucleic acid molecule, wherein said nucleic acid molecule encoding said nucleic acid-guided nuclease has at least 85% sequence identity to the nucleotide sequences of SEQ ID NO: 44 or SEQ ID NO: 24 and encodes an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 4;

(b) an engineered guide nucleic acid sequence capable of complexing with the nucleic acid-guided nuclease, wherein the engineered guide nucleic acid sequence is configured to hybridized to a TTTN sequence of a target region in a genome of a cell;

(c) an editing sequence having a change in sequence relative to the sequence of a target region in a genome of a cell;

wherein the nucleic acid-guided system results in a genome edit in the target region in the genome of the cell facilitated by the nucleic acid-guided nuclease, the engineered guide nucleic acid, and the editing sequence.

2. The nucleic acid-guided nuclease system of claim 1 , wherein the engineered guide nucleic acid sequence and the editing sequence are provided as a single nucleic acid.

3. The nucleic acid-guided nuclease system of claim 1 , wherein the nucleic acid molecule encoding the nucleic acid-guided nuclease is codon optimized for Escherichia Coli.

4. The nucleic acid-guided nuclease system of claim 1 , wherein the nucleic acid molecule encoding the nucleic acid-guided nuclease is codon optimized for Saccharomyces cerevisiae.

5. The nucleic acid-guided nuclease system of claim 1 , wherein the nucleic acid molecule encoding the nucleic acid-guided nuclease is codon optimized for mammalian cells.

6. The nucleic acid-guided nuclease system of claim 1 , wherein the target region is within a coding region of a protein.

7. The nucleic acid-guided nuclease system of claim 1 , wherein the target region is within a non-coding region of a protein.

8. The nucleic acid-guided nuclease system of claim 7 , wherein the target region is within a regulatory region of a protein.

9. The nucleic acid-guided nuclease system of claim 1 , wherein the editing sequence further comprises a mutation in a protospacer adjacent motif (PAM) sequence.

10. A method of modifying a target region in the genome of a cell, the method comprising:

(a) contacting a cell with the nucleic acid-guided nuclease system of claim 1 ; and

(b) allowing the nucleic-acid guided nuclease system to create a genome edit in a target region of the genome of the cell.

11. The method of claim 10 , wherein the engineered guide nucleic acid and the editing sequence are provided as a single nucleic acid.

12. The method of claim 10 , wherein the nucleic acid molecule encoding the nucleic acid-guided nuclease is codon optimized for Escherichia coli.

13. The method of claim 10 , wherein the nucleic acid molecule encoding the nucleic acid-guided nuclease is codon optimized for Saccharomyces cerevisiae.

14. The method of claim 10 , wherein the nucleic acid molecule encoding the nucleic acid-guided nuclease is codon optimized for mammalian cells.

15. The method of claim 10 , wherein the editing sequence further comprises a mutation in a protospacer adjacent motif (PAM) site.

16. The method of claim 10 , wherein the target region is within a eukaryotic cell.

17. The method of claim 10 , wherein the target region is within a bacterial cell.

18. The method of claim 10 , wherein the target region is within a plant cell.

19. The method of claim 10 , wherein the target region is within a mammalian cell.

Assignments (4)

Continuity (8)

References Cited (400)