Single-Nucleotide Microbial Genome Editing Using CRISPR-Cas12a
- Authors
- Lee, Ho Joung; Lee, Sang Jun
- Issue Date
- Mar-2024
- Publisher
- Humana Press Inc.
- Keywords
- 3′-truncated crRNA; Precise genome editing; Single-base; Cas12a
- Citation
- Methods in molecular biology (Clifton, N.J.), v.2760, pp 147 - 155
- Pages
- 9
- Journal Title
- Methods in molecular biology (Clifton, N.J.)
- Volume
- 2760
- Start Page
- 147
- End Page
- 155
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/73132
- DOI
- 10.1007/978-1-0716-3658-9_9
- ISSN
- 1940-6029
- Abstract
- Microbial genome editing can be achieved by donor DNA-directed mutagenesis and CRISPR-Cas12a-mediated negative selection. Single-nucleotide-level genome editing enables the manipulation of microbial cells exactly as designed. Here, we describe single-nucleotide substitutions/indels in the target DNA of E. coli genome using a mutagenic DNA oligonucleotide donor and truncated crRNA/Cas12a system. The maximal truncation of nucleotides at the 3'-end of the crRNA enables Cas12a-mediated single-nucleotide-level precise editing at galK targets in the genome of E. coli. © 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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Collections - College of Biotechnology & Natural Resource > Department of Systems Biotechnology > 1. Journal Articles
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