IP Library › Granted Patent US 12,454,684
Granted Patent B2
US 12,454,684 · App. 17/873,485 · Granted Oct 28, 2025

Methods using novel CPF1 mutations that enhance the DNA cleavage activity of

Inventors: Liyang Zhang (Coralville, IA); Christopher Anthony Vakulskas (North Liberty, IA); Nicole Mary Bode (Oxford, IA); Michael Allen Collingwood (North Liberty, IA); Kristin Renee Beltz (Cedar Rapids, IA); Mark Aaron Behlke (Coralville, IA)
Assignee: Integrated DNA Technologies, Inc.
C12N9/22C12N15/11C12N2310/20C12N2800/80
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Quick Facts
Patent No.
US 12,454,684
App. No.
17/873,485
Granted
Oct 28, 2025
Kind
B2
Abstract

The present disclosure concerns polynucleotides and amino acids of Acidaminococcus sp . Cas12a (Cpf1) and methods for their use for genome editing in eukaryotic cells.

Claims (16)

1. A method of performing genome editing at a target DNA site in an isolated eukaryotic cell in vitro or ex vivo using a CAS endonuclease system, comprising:

(a) introducing the CAS endonuclease system into the isolated eukaryotic cell in vitro or ex vivo, said CAS endonuclease system comprising:

(i) a Cas12a polypeptide encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 466, 467, 489, and 491; and

ii) a suitable guide RNA for the target DNA site; and

(b) contacting the target DNA site with the CAS endonuclease system.

2. A method of performing genome editing at a target DNA site in an isolated eukaryotic cell in vitro or ex vivo using a CAS endonuclease system, comprising:

(a) introducing the CAS endonuclease system into the isolated eukaryotic cell in vitro or ex vivo, said CAS endonuclease system comprising:

(i) a CRISPR-associated protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 465 and 493; and

(ii) a suitable guide RNA for the target DNA site; and

(b) contacting the target DNA site with the CAS endonuclease system.

3. The method according to claim 2 , wherein the CRISPR-associated protein comprises the amino acid sequence of SEQ ID NO: 465.

4. The method according to claim 2 , wherein the CRISPR-associated protein comprises the amino acid sequence of SEQ ID NO: 493.

5. The method according to claim 1 , wherein the Cas12a polypeptide is encoded by a nucleotide sequence that comprises SEQ ID NO: 466.

6. The method according to claim 1 , wherein the Cas12a polypeptide is encoded by a nucleotide sequence that comprises SEQ ID NO: 467.

7. The method according to claim 1 , wherein the Cas12a polypeptide is encoded by a nucleotide sequence that comprises SEQ ID NO: 489.

8. The method according to claim 1 , wherein the Cas12a polypeptide is encoded by a nucleotide sequence that comprises SEQ ID NO: 491.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 17, 2025
From: ZHANG, LIYANG; VAKULSKAS, CHRISTOPHER ANTHONY; BODE, NICOLE MARY; COLLINGWOOD, MICHAEL ALLEN; BELTZ, KRISTIN RENEE; BEHLKE, MARK AARON
To: INTEGRATED DNA TECHNOLOGIES, INC.
Reel/Frame 072285/0383 →
Continuity (6)
Continuation 16536256 · Aug 8, 2019
Provisional Application 62870268 · Jul 3, 2019
Provisional Application 62749607 · Oct 23, 2018
Provisional Application 62716138 · Aug 8, 2018
Related Publication 20230138679A1 · May 4, 2023
Related Publication 20250304932A9 · Oct 2, 2025
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