Highly efficient and specific regulation of gene expression using enhanced CRISPR-Cas12f system
- Authors
- Oh, Yeounsun; Gwon, Lee Wha; Lee, Hyomin K.; Hur, Junho K.; Park, Kwang-Hyun; Kim, Kee-Pyo; Lee, Seung Hwan
- Issue Date
- Jul-2024
- Publisher
- Springer Nature
- Citation
- Gene Therapy, v.31, no.7-8, pp 358 - 365
- Pages
- 8
- Journal Title
- Gene Therapy
- Volume
- 31
- Number
- 7-8
- Start Page
- 358
- End Page
- 365
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/75035
- DOI
- 10.1038/s41434-024-00458-w
- ISSN
- 0969-7128
1476-5462
- Abstract
- The recently developed CRISPR activator (CRISPRa) system uses a CRISPR-Cas effector-based transcriptional activator to effectively control the expression of target genes without causing DNA damage. However, existing CRISPRa systems based on Cas9/Cas12a necessitate improvement in terms of efficacy and accuracy due to limitations associated with the CRISPR-Cas module itself. To overcome these limitations and effectively and accurately regulate gene expression, we developed an efficient CRISPRa system based on the small CRISPR-Cas effector Candidatus Woesearchaeota Cas12f (CWCas12f). By engineering the CRISPR-Cas module, linking activation domains, and using various combinations of linkers and nuclear localization signal sequences, the optimized eCWCas12f-VPR system enabled effective and target-specific regulation of gene expression compared with that using the existing CRISPRa system. The eCWCas12f-VPR system developed in this study has substantial potential for controlling the transcription of endogenous genes in living organisms and serves as a foundation for future gene therapy and biological research. © The Author(s), under exclusive licence to Springer Nature Limited 2024.
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Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
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