Multiplex RNA-Guided Genome Engineering
US-2016168592-A1 · Jun 16, 2016 · US
Nucleic acid-guided nickases
US11965186B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-11965186-B2 |
| Application number | US-202217676218-A |
| Country | US |
| Kind code | B2 |
| Filing date | Feb 20, 2022 |
| Priority date | Jan 4, 2021 |
| Publication date | Apr 23, 2024 |
| Grant date | Apr 23, 2024 |
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- Title
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- Abstract
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Abstract
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The present disclosure provides engineered nucleic acid-guided nickases and optimized scaffolds for making rational, direct edits to nucleic acids in live cells.
First claim
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We claim: 1. A nucleic acid-guided nickase selected from the following nickases: MAD2017-H847A, having the amino acid sequence of SEQ ID NO: 5; and MAD2017-N870A, having the amino acid sequence of SEQ ID NO: 6. 2. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 5. 3. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 6. 4. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 5, in a nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 25, SEQ ID NO: 26, or SEQ ID NO: 27. 5. The nickase of claim 4 having the amino acid sequence of SEQ ID NO: 5, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 14. 6. The nickase of claim 4 having the amino acid sequence of SEQ ID NO: 5, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 15. 7. The nickase of claim 4 having the amino acid sequence of SEQ ID NO: 5, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 16. 8. The nickase of claim 4 having the amino acid sequence of SEQ ID NO: 5, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 25. 9. The nickase of claim 4 having the amino acid sequence of SEQ ID NO: 5, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 26. 10. The nickase of claim 4 having the amino acid sequence of SEQ ID NO: 5, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 27. 11. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 6, in a nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 25, SEQ ID NO: 26, or SEQ ID NO: 27. 12. The nickase of claim 11 having the amino acid sequence of SEQ ID NO: 6, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 14. 13. The nickase of claim 11 having the amino acid sequence of SEQ ID NO: 6, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 15. 14. The nickase of claim 11 having the amino acid sequence of SEQ ID NO: 6, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 16. 15. The nickase of claim 11 having the amino acid sequence of SEQ ID NO: 6, in the nucleic acid-guided nickase editing system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 25. 16. The nickase of claim 11 having the amino acid sequence of SEQ ID NO: 6, in the nucleic acid-guided nickase editing system with a gRNA scaffold having the nucleic acid sequence of SEQ ID NO: 26. 17. The nickase of claim 11 having the amino acid sequence of SEQ ID NO: 6, in the system with a gRNA scaffold having a nucleic acid sequence of SEQ ID NO: 27. 18. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 5, in a nucleic acid-guided nickase editing system with a native CRISPR repeat having a nucleic acid sequence of SEQ ID NO: 12 and a native tracr RNA having a nucleic acid sequence of SEQ ID NO: 13. 19. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 6, in a nucleic acid-guided nickase editing system with a native CRISPR repeat having a nucleic acid sequence of SEQ ID NO: 12 and a native tracr RNA having a nucleic acid sequence of SEQ ID NO: 13. 20. The nickase of claim 1 having the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO; 4, in a nucleic acid-guided nickase editing system comprising a guide RNA wherein the guide comprises from 5′ to 3′ a guide sequence, a homology region and SEQ ID NO. 30.
Assignees
Inventors
Classifications
- C12N9/22Primary
Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title
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} · CPC title
in mammalian cells · CPC title
Type II site-specific deoxyribonuclease (3.1.21.4) · CPC title
involving clustered regularly interspaced short palindromic repeats [CRISPR] · CPC title
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Frequently asked questions
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- What does patent US11965186B2 cover?
- The present disclosure provides engineered nucleic acid-guided nickases and optimized scaffolds for making rational, direct edits to nucleic acids in live cells.
- Who is the assignee on this patent?
- Inscripta Inc
- What technology area does this patent fall under?
- Primary CPC classification C12N9/22. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Apr 23 2024 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 12 related publications on this page (citations in our corpus or others sharing the same primary CPC).