MagnEdit-interacting Factors That Recruit DNA-editing Enzymes to Single Base Targets

Overview

Journal Life Sci Alliance

Specialties Biochemistry
Biology
Cell Biology
Molecular Biology

Date 2020 Feb 26

PMID 32094150

Citations 6

Authors

Jennifer L McCann

Daniel J Salamango

Emily K Law

William L Brown

Reuben S Harris

Affiliations

Soon will be listed here.

Abstract

Although CRISPR/Cas9 technology has created a renaissance in genome engineering, particularly for gene knockout generation, methods to introduce precise single base changes are also highly desirable. The covalent fusion of a DNA-editing enzyme such as APOBEC to a Cas9 nickase complex has heightened hopes for such precision genome engineering. However, current cytosine base editors are prone to undesirable off-target mutations, including, most frequently, target-adjacent mutations. Here, we report a method to "attract" the DNA deaminase, APOBEC3B, to a target cytosine base for specific editing with minimal damage to adjacent cytosine bases. The key to this system is fusing an APOBEC-interacting protein (not APOBEC itself) to Cas9n, which attracts nuclear APOBEC3B transiently to the target site for editing. Several APOBEC3B interactors were tested and one, hnRNPUL1, demonstrated proof-of-concept with successful C-to-T editing of episomal and chromosomal substrates and lower frequencies of target-adjacent events.

Citing Articles

Structure-guided inhibition of the cancer DNA-mutating enzyme APOBEC3A.

Harjes S, Kurup H, Rieffer A, Bayarjargal M, Filitcheva J, Su Y Nat Commun. 2023; 14(1):6382.

PMID: 37821454 PMC: 10567711. DOI: 10.1038/s41467-023-42174-w.

APOBEC Reporter Systems for Evaluating diNucleotide Editing Levels.

Rieffer A, Chen Y, Salamango D, Moraes S, Harris R CRISPR J. 2023; 6(5):430-446.

PMID: 37672599 PMC: 10611974. DOI: 10.1089/crispr.2023.0027.

Structure-guided inhibition of the cancer DNA-mutating enzyme APOBEC3A.

Harjes S, Kurup H, Rieffer A, Bayaijargal M, Filitcheva J, Su Y bioRxiv. 2023; .

PMID: 36824964 PMC: 9949147. DOI: 10.1101/2023.02.17.528918.

Inhibition of base editors with anti-deaminases derived from viruses.

Liu Z, Chen S, Lai L, Li Z Nat Commun. 2022; 13(1):597.

PMID: 35105899 PMC: 8807840. DOI: 10.1038/s41467-022-28300-0.

Approaches to Enhance Precise CRISPR/Cas9-Mediated Genome Editing.

Denes C, Cole A, Aksoy Y, Li G, Neely G, Hesselson D Int J Mol Sci. 2021; 22(16).

PMID: 34445274 PMC: 8395304. DOI: 10.3390/ijms22168571.

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