Efficient CRISPR-Cas12f1-Mediated Multiplex Bacterial Genome Editing via Low-Temperature Recovery.Efficient CRISPR-Cas12f1-Mediated Multiplex Bacterial Genome Editing via Low-Temperature Recovery
- Authors
- Lim, Se Ra; Kim, Hyun Ju; Lee, Sang Jun
- Issue Date
- Jul-2024
- Publisher
- 한국미생물·생명공학회
- Keywords
- AsCas12f1; Nucleotide editing; multiple targets; recovery temperature; truncated guide RNA
- Citation
- Journal of microbiology and biotechnology, v.34, no.7, pp 1 - 8
- Pages
- 8
- Journal Title
- Journal of microbiology and biotechnology
- Volume
- 34
- Number
- 7
- Start Page
- 1
- End Page
- 8
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/74542
- DOI
- 10.4014/jmb.2403.03033
- ISSN
- 1017-7825
1738-8872
- Abstract
- CRISPR-Cas system is being used as a powerful genome editing tool with developments focused on enhancing its efficiency and accuracy. Recently, the miniature CRISPR-Cas12f1 system, which is small enough to be easily loaded onto various vectors for cellular delivery, has gained attention. In this study, we explored the influence of temperature conditions on multiplex genome editing using CRISPR-Cas12f1 in an Escherichia coli model. It was revealed that when two distinct targets in the genome are edited simultaneously, the editing efficiency can be enhanced by allowing cells to recover at a reduced temperature during the editing process. Additionally, employing 3'-end truncated sgRNAs facilitated the simultaneous single-nucleotide level editing of three targets. Our results underscore the potential of optimizing recovery temperature and sgRNA design protocols in developing more effective and precise strategies for multiplex genome editing across various organisms.
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Collections - College of Biotechnology & Natural Resource > Department of Systems Biotechnology > 1. Journal Articles
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