IP Library › Granted Patent US 12,264,342
Granted Patent B2
US 12,264,342 · App. 17/430,013 · Granted Apr 1, 2025

Patent

Inventors: Sarah Franz Beaudoin (Iowa City, IA); Michael Allen Collingwood (North Liberty, IA); Christopher Anthony Vakulskas (North Liberty, IA)
Assignee: INTEGRATED DNA TECHNOLOGIES, INC.
C12N9/22C12N15/11C12N2310/20
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Quick Facts
Patent No.
US 12,264,342
App. No.
17/430,013
Granted
Apr 1, 2025
Kind
B2
Abstract

This invention pertains to mutant Lachnospiraceae bacterium ND2006 (Lb) Cas12a nucleic acids and proteins for use in CRISPR/Cas12a endonuclease systems, and their methods of use. In particular, the invention pertains to an isolated mutant LbCas12a protein, wherein the isolated mutant LbCas12a protein is active in a CRISPR/Cas12a endonuclease system. The invention also includes isolated nucleic acids encoding mutant LbCas12a proteins, ribonucleoprotein complexes and CRISPR/Cas12a endonuclease systems having mutant LbCas12a proteins.

Claims (12)

1. An isolated mutant LbCas12a protein comprising

a single amino acid substitution mutation introduced into the wild-type LbCas12a protein comprising E795L (SEQ ID NO: 5).

2. An isolated ribonucleoprotein complex, comprising:

(a) the mutant LbCas12a protein of claim 1 ; and

(b) a gRNA complex;

wherein the isolated ribonucleoprotein complex is active as a CRISPR/Cas12a endonuclease system, wherein the resultant CRISPR/Cas12a endonuclease system displays maintained on-target editing activity relative to a wild-type CRISPR/Cas12a endonuclease system.

3. A CRISPR/Cas12a endonuclease system comprising a mutant LbCas12a protein and a gRNA, wherein the CRISPR/Cas12a endonuclease system displays maintained on-target editing activity relative to a wild-type CRISPR/Cas12a endonuclease system, and wherein the mutant LbCas12a protein comprises a single amino acid substitution mutation introduced into the wild-type LbCas12a protein comprising E795L (SEQ ID NO: 5).

4. The CRISPR/Cas12a endonuclease system of claim 3 , wherein the CRISPR/Cas12a endonuclease system is encoded by a DNA expression vector.

5. The CRISPR/Cas12a endonuclease system of claim 4 , wherein the DNA expression vector comprises a plasmid-borne vector.

6. The CRISPR/Cas12a endonuclease system of claim 5 , wherein the DNA expression vector is selected from a bacterial expression vector and a eukaryotic expression vector.

7. An isolated nucleic acid encoding a mutant LbCas12a protein, wherein the mutant LbCas12a protein is active in a CRISPR/Cas12a endonuclease system, wherein the CRISPR/Cas12a endonuclease system displays maintained on-target editing activity relative to a wild-type CRISPR/Cas12a endonuclease system, and wherein the mutant LbCas12a protein comprises a single amino acid substitution mutation introduced into the wild-type LbCas12a protein comprising E795L (SEQ ID NO: 5).

8. The isolated nucleic acid encoding a mutant LbCas12a protein of claim 7 , wherein the isolated nucleic acid encoding the mutant LbCas12a protein is SEQ ID NO: 12.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 30, 2021
From: BEAUDOIN, SARAH FRANZ; COLLINGWOOD, MICHAEL ALLEN; VAKULSKAS, CHRISTOPHER ANTHONY
To: INTEGRATED DNA TECHNOLOGIES, INC.
Reel/Frame 057658/0405 →
Continuity (2)
Provisional Application 62808984 · Feb 22, 2019
Related Publication 20230040148A1 · Feb 9, 2023
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