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Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing
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 c...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9287484/ https://www.ncbi.nlm.nih.gov/pubmed/35671823 http://dx.doi.org/10.1016/j.jbc.2022.102103 |
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author | Ma, Lixia Xing, Jiani Li, Qian Zhang, Zhiying Xu, Kun |
author_facet | Ma, Lixia Xing, Jiani Li, Qian Zhang, Zhiying Xu, Kun |
author_sort | Ma, Lixia |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9287484 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-92874842022-07-19 Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing Ma, Lixia Xing, Jiani Li, Qian Zhang, Zhiying Xu, Kun J Biol Chem Research Article 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. American Society for Biochemistry and Molecular Biology 2022-06-06 /pmc/articles/PMC9287484/ /pubmed/35671823 http://dx.doi.org/10.1016/j.jbc.2022.102103 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Ma, Lixia Xing, Jiani Li, Qian Zhang, Zhiying Xu, Kun Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
title | Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
title_full | Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
title_fullStr | Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
title_full_unstemmed | Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
title_short | Development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
title_sort | development of a universal antibiotic resistance screening reporter for improving efficiency of cytosine and adenine base editing |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9287484/ https://www.ncbi.nlm.nih.gov/pubmed/35671823 http://dx.doi.org/10.1016/j.jbc.2022.102103 |
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