Improved Cas12a Proteins for Accurate and Efficient Genome Editing
Patent Status
| Country | Type | Number | Dated | Case |
| United States Of America | Published Application | 20220315914 | 10/06/2022 | 2019-162 |
Brief Description
Mutated versions of Cas12a that remove its non-specific ssDNA cleavage activity without affecting site-specific double-stranded DNA cutting activity. These mutant proteins, in which a short amino acid sequence is deleted or changed, provide improved genome editing tools that will avoid potential off-target editing due to random ssDNA nicking.
Suggested uses
Genome editing in animals, plants, and human cells.
Advantages
Accurate and efficient genome editing.
Background: Cas12a (formerly called Cpf1) is a type V CRISPR-Cas enzyme derived from bacteria that is used for RNA-guided genome editing in animal, plant and human cells. However, Cas12a possesses an additional enzymatic activity in which a DNA target-bound Cas12a can rapidly and non-specifically degrade any single-stranded DNA (ssDNA) substrate in a sequence-independent manner. This enzymatic activity is endonucleolytic, which means that the ssDNA substrate does not need a free 5' or 3' end to be cut. For this reason, natural Cas12a-type enzymes have the potential to induce off-target genome editing due to nicking of exposed ssDNA in cells.
Contact
- Terri Sale
- terri.sale@berkeley.edu
- tel: View Phone Number.
Inventors
- Doudna, Jennifer A.
Other Information
Keywords
genome editing, Cas12a
Additional Technologies by these Inventors
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- Genome Editing via LNP-Based Delivery of Efficient and Stable CRISPR-Cas Editors
- Type III CRISPR-Cas System for Robust RNA Knockdown and Imaging in Eukaryotes
- Cas12-mediated DNA Detection Reporter Molecules
- Highly Multiplexed Tagging Methods for RNA Imaging and Other Applications
- Improved guide RNA and Protein Design for CasX-based Gene Editing Platform
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- Miniature Type VI CRISPR-Cas Systems and Methods of Use
- RNA-directed Cleavage and Modification of DNA using CasY (CRISPR-CasY)
- CasX Nickase Designs, Tans Cleavage Designs & Structure
- In Vivo Gene Editing Of Tau Locus Via Liponanoparticle Delivery
- Methods and Compositions for Modifying a single stranded Target Nucleic Acid
- A Dual-RNA Guided CasZ Gene Editing Technology
- Single-Stranded Nucleic Acid Detection And Imaging System Using Cas9
- CRISPR-CAS EFFECTOR POLYPEPTIDES AND METHODS OF USE THEREOF (“Cas-VariPhi”)
- A Protein Inhibitor Of Cas9
- RNA-directed Cleavage and Modification of DNA using CasX (CRISPR-CasX)
- Compositions and Methods for Genome Editing
- Split-Cas9 For Regulatable Genome Engineering
- Methods to Interfere with Prokaryotic and Phage Translation and Noncoding RNA
- Minimal RNA Targeting CRISPR Cas Systems
- Variant Cas12a Protein Compositions and Methods of Use
- CRISPR CASY COMPOSITIONS AND METHODS OF USE
- Single Conjugative Vector for Genome Editing by RNA-guided Transposition
- CRISPR-CAS EFFECTOR POLYPEPTIDES AND METHODS OF USE THEREOF
- Engineered/Variant Hyperactive CRISPR CasPhi Enzymes And Methods Of Use Thereof
- Methods Of Use Of Cas12L/CasLambda In Plants
- Type V CRISPR/CAS Effector Proteins for Cleaving ssDNA and Detecting Target DNA
- THERMOSTABLE RNA-GUIDED ENDONUCLEASES AND METHODS OF USE THEREOF (GeoCas9)
- Structure-Guided Methods Of Cas9-Mediated Genome Engineering
- Efficient Site-Specific Integration Of New Genetic Information Into Human Cells
- CRISPR-Cas Effector Polypeptides and Methods of Use Thereof
- Virus-encoded DNA-binding Proteins
- Class 2 CRISPR/Cas COMPOSITIONS AND METHODS OF USE
- Compositions and Methods of Use for Variant Csy4 Endoribonucleases
- Methods and Compositions for Controlling Gene Expression by RNA Processing
