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PreprintCited by 19CRISPR and Genetic EngineeringRNA regulation and diseaseBacterial Genetics and Biotechnology

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Engineered minimal type I CRISPR-Cas system for transcriptional activation and base editing in human cells

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Guo, Jing, Gong, Luyao, Yu, Haiying, Li, Ming, Liu, Zhenquan, Fan, Shuru, Yang, Changjialian, Zhao, Dahe, Han, Jing, Xiang, Hua·2024·Nature Communications
DOI 10.1101/2024.01.26.577312PMC PMC11343773

Preprint — published as 10.1038/s41467-024-51695-x

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Abstract

Type I CRISPR-Cas systems are widespread and have exhibited remarkable versatility and efficiency in genome editing and gene regulation in prokaryotes. However, due to the multi-subunit composition and large size, their application in eukaryotes has not been thoroughly investigated. Here, we demonstrate that the type I-F2 Cascade, the most compact among type I systems and significantly smaller than SpCas9, can be developed into programmable tools for use in human cells. For transcriptional activation, the efficiency of the tool based on the engineered I-F2 system can match or surpass that of dCas9. Besides, narrow editing windows limit the application of base editors. Although the R-loop formed by Cascade is much wider than that by Cas9 or Cas12, the potential of base editing with Cascade has not yet been explored. We successfully created a base editor with the I-F2 Cascade, which induces a considerably wide editing window (∼30 nt) with a bimodal distribution. The wide editing window can expand the range of targetable sites and can be useful for disrupting functional sequences and genetic screening. The editing efficiency can achieve 50% in human cells. This research underscores the application potential of compact type I systems in eukaryotes and developed a new base editor with an extraordinary wide editing window.

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Metadata

Authors
Guo, J., Gong, L., Yu, H., Li, M., Liu, Z., Fan, S., Yang, C., Zhao, D., Han, J., & Xiang, H.
Affiliations
Chinese Academy of Sciences (CN)
Corresponding author
  • Gong, Luyao
  • Xiang, Hua
ORCID
Venue
Nature Communications
Publisher
openRxiv
Published
2024-01-26
Volume
15
Issue
1
Pages
7277-7277
ISSN
2041-1723 (ISSN-L)
DOI
10.1101/2024.01.26.577312
PMC
PMC11343773
OpenAlex
W4401808393
Cited by
19
References
1
Citations by year
2026: 5 · 2025: 11 · 2024: 3
FWCI
2.58
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (2020YFA0906800, 32100499, 32150020, 32230061, XDA24020101)
  • Chinese Academy of Sciences (XDA24020101)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
type I CRISPR-Cas, transcriptional activation, base editing, human cells, genome editing, gene expression regulation
MeSH terms
Gene Editing, CRISPR-Associated Protein 9, RNA, Guide, CRISPR-Cas Systems, Genetic Engineering, Humans, Transcriptional Activation, HEK293 Cells, CRISPR-Cas Systems

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