IP Library › Granted Patent US 12,054,707
Granted Patent B2
US 12,054,707 · App. 17/227,799 · Granted Aug 6, 2024

High fidelity restriction endonucleases

Inventors: Zhenyu Zhu (Beverly, MA); Aine Quimby (Newton, NH); Shuang-Yong Xu (Lexington, MA); Shengxi Guan (Stoneham, MA); Hua Wei (Ipswich, MA); Penghua Zhang (Lexington, MA); Dapeng Sun (Arlington, MA); Siu-hong Chan (Ipswich, MA)
Assignee: New England Biolabs, Inc.
C12N15/1058C12N9/16C12N9/22C12N15/1034C12N15/1086
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Quick Facts
Patent No.
US 12,054,707
App. No.
17/227,799
Granted
Aug 6, 2024
Kind
B2
Abstract

Compositions and methods are provided for enzymes with altered properties that involve a systematic approach to mutagenesis and a screening assay that permits selection of the desired proteins. Embodiments of the method are particularly suited for modifying specific properties of restriction endonucleases such as star activity. The compositions includes restriction endonucleases with reduced star activity as defined by an overall fidelity index improvement factor.

Claims (14)

1. A composition comprising a variant BspQI restriction endonuclease, wherein the variant BspQI restriction endonuclease comprises an amino acid sequence having one, two, or all three of the following substitutions:

a substitution at a position corresponding to 244 in SEQ ID NO: 16;

a substitution at a position corresponding to 279 in SEQ ID NO: 16; and

a substitution at a position corresponding to 388 in SEQ ID NO: 16.

2. A composition according to claim 1 , wherein the substitution corresponding to position 244 is D244A.

3. A composition according to claim 1 , wherein the substitution corresponding to position 279 is K279A, K279D, K279E, K279F, K279L, K279P, or K279Y.

4. A composition according to claim 1 , wherein the substitution corresponding to position 279 is K279P.

5. A composition according to claim 1 , wherein the substitution corresponding to position 388 is R388A, R388D, R388E, R388F, R388L, R388P, or R388Y.

6. A composition according to claim 1 , wherein the substitution corresponding to position 388 is R388F.

7. A composition according to claim 1 , wherein the variant BspQI restriction endonuclease comprises an amino acid sequence having the following substitutions:

a substitution at a position corresponding to 279 in SEQ ID NO: 16; and

a substitution at a position corresponding to 388 in SEQ ID NO: 16.

8. The composition according to claim 7 , wherein the substitutions at positions 279 and 388, are K279A and R388A.

9. The composition according to claim 7 , wherein the substitutions at positions 279 and 388, are K279P and R388F.

Assignments (2)
NOTICE OF GRANT OF SECURITY INTEREST IN PATENTS Recorded Sep 27, 2023
From: NEW ENGLAND BIOLABS, INC.
To: BANK OF AMERICA, N.A., AS ADMINISTRATIVE AGENT
Reel/Frame 065044/0729 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 19, 2022
From: ZHENYU, ZHU; QUIMBY, AINE; XU, SHUANG-YONG; GUAN, SHENGXI; WEI, HUA; ZHANG, PENGHUA; CHAN, SIU-HONG
To: NEW ENGLAND BIOLABS, INC.
Reel/Frame 061136/0034 →