CRISPR 간섭에 필요한 sgRNA 표적 인식 서열 길이의 결정Determination of the Length of Target Recognition Sequence in sgRNA Required for CRISPR Interference
- Authors
- Kim, Bumjoon; Kim, Byeong Chan; Lee, Ho Joung; Lee, Sang Jun
- Issue Date
- Dec-2021
- Publisher
- Korean Society for Microbiolog and Biotechnology
- Keywords
- CRISPR interference; D-galactose; Galpromoter; Single-molecular guide RNA
- Citation
- Microbiology and Biotechnology Letters, v.49, no.4, pp 534 - 542
- Pages
- 9
- Journal Title
- Microbiology and Biotechnology Letters
- Volume
- 49
- Number
- 4
- Start Page
- 534
- End Page
- 542
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/54953
- DOI
- 10.48022/mbl.2111.11003
- ISSN
- 1598-642X
- Abstract
- Single-molecular guide RNA (sgRNA) plays a role in recognizing the DNA target sequence in CRISPR technology for genome editing and gene expression control. In this study, we systematically compared the length of the target recognition sequence in sgRNAs required for genome editing using Cas9-NG (an engineered Cas9 recognizing 5’-NG as PAM sequence) and gene expression control using deactivated Cas9-NG (dCas9-NG) by targeting the gal promoter in E. coli. In the case of genome editing, the truncation of three nucleotides in the target recognition sequence (TRS) of sgRNA was allowed. In gene expression regulation, we observed that target recognition and binding were possible even if eleven nucleotides were deleted from twenty nucleotides of the TRS. When 4 or more nucleotides are truncated in the TRS of the sgRNA, it is thought that the sgRNA/Cas9-NG complex can specifically bind to the target DNA sequence, but lacks endonuclease activity to perform genome editing. Our study will be helpful in the development of artificial transcription factors and various CRISPR technologies in the field of synthetic biology. © 2021, The Korean Society for Microbiology and Biotechnology
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Collections - College of Biotechnology & Natural Resource > Department of Systems Biotechnology > 1. Journal Articles
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