Multiplex RNA-Guided Genome Engineering
US-2016168592-A1 · Jun 16, 2016 · US
Engineered enzyme
US12146170B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-12146170-B2 |
| Application number | US-202117551391-A |
| Country | US |
| Kind code | B2 |
| Filing date | Dec 15, 2021 |
| Priority date | Oct 22, 2018 |
| Publication date | Nov 19, 2024 |
| Grant date | Nov 19, 2024 |
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Abstract
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The present disclosure provides a novel catalytically inactive MAD7 nuclease (dMAD7) that retains the ability to bind DNA in a sequence-specific manner. The MAD7 nuclease from which the dMAD7 has been derived was isolated from Eubacterium rectale.
First claim
Opening claim text (preview).
We claim: 1. A catalytically inactive CRISPR nuclease comprising a variant amino acid sequence of SEQ ID NO: 1, and wherein the variant amino acid sequence comprises an alanine residue at amino acid position 877 as compared to SEQ ID NO: 1. 2. The nuclease of claim 1 , wherein the nuclease binds a bacteria genome in a sequence-specific manner. 3. The nuclease of claim 1 , wherein the nuclease binds a yeast genome in a sequence-specific manner. 4. The nuclease of claim 1 , wherein the nuclease binds a mammalian genome in a sequence specific manner. 5. A vector comprising a nucleic acid sequence encoding the nuclease of claim 1 . 6. The vector of claim 5 , wherein the vector further comprises a nucleic acid sequence encoding a guide RNA. 7. The vector of claim 5 , wherein the vector further comprises a nucleic acid sequence encoding a donor nucleic acid. 8. The vector of claim 5 , wherein the nucleic acid sequence encoding the nuclease further comprises a nucleic acid sequence encoding one or more nuclear localization sequences. 9. The vector of claim 5 , wherein the nucleic acid sequence encoding the nuclease is operably linked to a promoter. 10. The vector of claim 9 , wherein the promoter is an inducible promoter. 11. The vector of claim 9 , wherein the promoter is a constitutive promoter. 12. The vector of claim 6 , wherein the nucleic acid sequence encoding a guide RNA is operably linked to a promoter. 13. The vector of claim 12 , wherein the promoter is an inducible promoter. 14. The vector of claim 12 , wherein the promoter is a constitutive promoter. 15. The vector of claim 9 , wherein the nucleic acid sequence encoding the nuclease is codon-optimized for expression in a eukaryotic cell. 16. The vector of claim 15 , wherein the eukaryotic cell is a fungal cell. 17. The vector of claim 16 , wherein the fungal cell is a yeast cell. 18. The vector of claim 15 , wherein the eukaryotic cell is a human cell. 19. The vector of claim 15 , wherein the eukaryotic cell is a plant cell. 20. The vector of claim 9 , wherein the nucleic acid sequence encoding the nuclease is codon-optimized for expression in a prokaryotic cell.
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Classifications
- C12N9/22Primary
Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title
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- What does patent US12146170B2 cover?
- The present disclosure provides a novel catalytically inactive MAD7 nuclease (dMAD7) that retains the ability to bind DNA in a sequence-specific manner. The MAD7 nuclease from which the dMAD7 has been derived was isolated from Eubacterium rectale.
- 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 Nov 19 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).