IP Library › Granted Patent US 12,297,467
Granted Patent B2
US 12,297,467 · App. 17/846,771 · Granted May 13, 2025

Lipid-encapsulated dual-cleaving endonuclease for DNA and gene editing

Inventors: David R. Edgell (London, CA); Thomas A. McMurrough (London, CA); Brent E. Stead (Toronto, CA); Odisho K. Israel (Toronto, CA)
Assignee: SPECIFIC BIOLOGICS INC.
C12N9/22A61K31/7088A61K38/465C12N15/11C12N15/907C12N2310/20C12N2800/80
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Quick Facts
Patent No.
US 12,297,467
App. No.
17/846,771
Granted
May 13, 2025
Kind
B2
Abstract

Methods to edit genes by administering a chimeric nuclease to a cell or organism without the use of a viral vector.

Claims (18)

1. A nuclease comprising a modified Staphylococcus aureus Cas9, wherein said modified Staphylococcus aureus Cas9 comprises an aspartic acid to glutamic acid substitution at an amino acid corresponding to position 10 of SEQ ID NO: 13, and wherein the amino acid sequence of the Staphylococcus aureus Cas9 has at least 99% sequence identity to SEQ ID NO: 13.

2. The nuclease of claim 1 , wherein the modified Staphylococcus aureus Cas9 comprises a histidine to alanine substitution at an amino acid corresponding to position 557 of SEQ ID NO: 13.

3. The nuclease of claim 1 , further comprising an I-Tevl nuclease domain.

4. The nuclease of claim 3 , wherein the I-Tevl nuclease domain comprises the amino acid sequence set forth in SEQ ID NO: 6.

5. The nuclease of claim 3 , wherein the I-Tevl nuclease domain comprises a substitution or deletion of the methionine at position 1 of SEQ ID NO: 6, and otherwise comprises the amino acid sequence set forth in SEQ ID NO: 6.

6. The nuclease of claim 3 , further comprising a linker coupling the modified Staphylococcus aureus Cas9 and the I-Tevl nuclease domain.

7. The nuclease of claim 6 , wherein the linker comprises an amino acid sequence set forth in any one or more of SEQ ID NO: 7 to SEQ ID NO 12.

8. The nuclease of claim 1 , bound to a guide RNA.

9. The nuclease of claim 8 , wherein the guide RNA comprises one or more bridged nucleic acids.

10. A chimeric nuclease comprising an I-Tevl nuclease domain, a linker, a Staphylococcus aureus Cas9, and a guide RNA, wherein the Staphylococcus aureus Cas9 comprises an aspartic acid to glutamic acid substitution at an amino acid corresponding to position 10 of SEQ ID NO: 13, and wherein the amino acid sequence of the Staphylococcus aureus Cas9 has at least 99% sequence identity to SEQ ID NO: 13.

11. The chimeric nuclease of claim 10 , wherein the Staphylococcus aureus Cas9 comprises a histidine to alanine substitution at an amino acid corresponding to position SS7 of SEQ ID NO: 13.

12. The chimeric nuclease of claim 10 , wherein the I-Tevl nuclease domain comprises the amino acid sequence set forth in SEQ ID NO: 6.

13. The chimeric nuclease of claim 10 , wherein the I-Tevl nuclease domain comprises a substitution or deletion of the methionine at position 1 of SEQ ID NO: 6, and otherwise comprises the amino acid sequence set forth in SEQ ID NO: 6.

14. The chimeric nuclease of claim 10 , wherein the linker comprises an amino acid sequence set forth in any one or more of SEQ ID NO: 7 to SEQ ID NO 12.

15. A formulation comprising the chimeric nuclease of claim 10 and a lipid nanoparticle.

16. A method to genetically modify the genome of a cell, the method comprising contacting the cell with the chimeric nuclease of claim 11 .

17. A nucleic acid encoding the nuclease of claim 1 .

18. A nucleic acid encoding the chimeric nuclease of claim 10 .

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 21, 2023
From: EDGELL, DAVID R.; MCMURROUGH, THOMAS A.; STEAD, BRENT E.; ISRAEL, ODISHO K.
To: SPECIFIC BIOLOGICS INC.
Reel/Frame 064652/0108 →
Continuity (5)
Continuation 17514585 · Oct 29, 2021
Continuation PCTIB2020054229 · May 4, 2020
Provisional Application 62842586 · May 3, 2019
Provisional Application 63019423 · May 3, 2020
Related Publication 20230016280A1 · Jan 19, 2023
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Cited By (1)
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