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Cas12a Base Editors Induce Efficient and Specific Editing with Low DNA Damage Response.
- Source :
-
Cell reports [Cell Rep] 2020 Jun 02; Vol. 31 (9), pp. 107723. - Publication Year :
- 2020
-
Abstract
- The advent of base editors (BEs) holds great potential for correcting pathogenic-related point mutations to treat relevant diseases. However, Cas9 nickase (nCas9)-derived BEs lead to DNA double-strand breaks, which can trigger unwanted DNA damage response (DDR). Here, we show that the original version of catalytically dead Cas12a (dCas12a)-conjugated BEs induce a basal level of DNA breaks and minimally activate DDR proteins, including H2AX, ATM, ATR, and p53. By fusing dCas12a with engineered human apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3A (APOBEC3A), we further develop the BEACON (base editing induced by human APOBEC3A and Cas12a without DNA break) system to achieve enhanced deamination efficiency and editing specificity. Efficient C-to-T editing is achieved by BEACON in mammalian cells at levels comparable to AncBE4max, with only low levels of DDR and minimal RNA off-target mutations. Importantly, BEACON induces in vivo base editing in mouse embryos, and targeted C-to-T conversions are detected in F0 mice.<br />Competing Interests: Declaration of Interests J.C., L.Y., X.H., B.Y., X.W., C.D., and W.Y. have filed patent applications on aspects on this work. The authors declare no competing non-financial interests.<br /> (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Subjects :
- 17-Hydroxysteroid Dehydrogenases genetics
Animals
Ataxia Telangiectasia Mutated Proteins metabolism
Bacterial Proteins genetics
CRISPR-Associated Proteins genetics
Cytidine metabolism
Cytidine Deaminase genetics
Cytidine Deaminase metabolism
DNA Replication
Deamination
Endodeoxyribonucleases genetics
Female
HEK293 Cells
Humans
Mice
Mice, Inbred C57BL
Mice, Inbred ICR
Phosphorylation
Proteins genetics
Proteins metabolism
Thymidine metabolism
Tumor Suppressor Protein p53 metabolism
Ubiquitins metabolism
Bacterial Proteins metabolism
CRISPR-Associated Proteins metabolism
DNA Damage
Endodeoxyribonucleases metabolism
Gene Editing methods
Subjects
Details
- Language :
- English
- ISSN :
- 2211-1247
- Volume :
- 31
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Cell reports
- Publication Type :
- Academic Journal
- Accession number :
- 32492431
- Full Text :
- https://doi.org/10.1016/j.celrep.2020.107723