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Tytuł pozycji:

Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing.

Tytuł:
Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing.
Autorzy:
Ma L; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China; Central Laboratory, Changzhi Medical College, Changzhi, Shanxi, China.
Xing J; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
Li Q; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China.
Zhang Z; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China. Electronic address: .
Xu K; Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China. Electronic address: .
Źródło:
The Journal of biological chemistry [J Biol Chem] 2022 Jul; Vol. 298 (7), pp. 102103. Date of Electronic Publication: 2022 Jun 06.
Typ publikacji:
Journal Article
Język:
English
Imprint Name(s):
Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology
MeSH Terms:
Adenine*/chemistry
Cytosine*/chemistry
Drug Resistance, Microbial*
Gene Editing*/methods
Microbial Sensitivity Tests*/methods
Contributed Indexing:
Keywords: adenine base editor; base-editing activity; cytosine base editor; screening; universal reporter
Substance Nomenclature:
8J337D1HZY (Cytosine)
JAC85A2161 (Adenine)
Entry Date(s):
Date Created: 20220607 Date Completed: 20220726 Latest Revision: 20220728
Update Code:
20240105
PubMed Central ID:
PMC9287484
DOI:
10.1016/j.jbc.2022.102103
PMID:
35671823
Czasopismo naukowe
Base editing has emerged as a revolutionary technology for single nucleotide modifications. The cytosine and adenine base editors (CBEs and ABEs) have demonstrated great potential in clinical and fundamental research. However, screening and isolating target-edited cells remains challenging. In the current study, we developed a universal Adenine and Cytosine Base-Editing Antibiotic Resistance Screening Reporter (ACBE-ARSR) for improving the editing efficiency. To develop the reporter, the CBE-ARSR was first constructed and shown to be capable of enriching cells for those that had undergone CBE editing activity. Then, the ACBE-ARSR was constructed and was further validated in the editing assays by four different CBEs and two versions of ABE at several different genomic loci. Our results demonstrated that ACBE-ARSR, compared to the reporter of transfection (RoT) screening strategy, improved the editing efficiency of CBE and ABE by 4.6- and 1.9-fold on average, respectively. We found the highest CBE and ABE editing efficiencies as enriched by ACBE-ARSR reached 90% and 88.7%. Moreover, we also demonstrated ACBE-ARSR could be employed for enhancing simultaneous multiplexed genome editing. In conclusion, both CBE and ABE activity can be improved significantly using our novel ACBE-ARSR screening strategy, which we believe will facilitate the development of base editors and their application in biomedical and fundamental research studies.
Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.
(Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

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