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Cited by 24CRISPR and Genetic EngineeringBiosensors and Analytical DetectionAdvanced biosensing and bioanalysis techniques

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Repurposing Type I-A CRISPR-Cas3 for a robust diagnosis of human papillomavirus (HPV)

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Hu, Tao, Ji, Quanquan, Ke, Xinxin, Zhou, Hufeng, Zhang, Senfeng, Ma, Shengsheng, Yu, Chenlin, Ju, Wenjun, Lu, Meiling, Lin, Yu, Ou, Yangjing, Zhou, Yingsi, Xiao, Yibei, Xu, Chunlong, Hu, Chunyi·2024·Communications Biology
DOI 10.1038/s42003-024-06537-3PMC PMC11246428

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Abstract

R-loop-triggered collateral single-stranded DNA (ssDNA) nuclease activity within Class 1 Type I CRISPR-Cas systems holds immense potential for nucleic acid detection. However, the hyperactive ssDNase activity of Cas3 introduces unwanted noise and false-positive results. In this study, we identified a novel Type I-A Cas3 variant derived from Thermococcus siculi , which remains in an auto-inhibited state until it is triggered by Cascade complex and R-loop formation. This Type I-A CRISPR-Cas3 system not only exhibits an expanded protospacer adjacent motif (PAM) recognition capability but also demonstrates remarkable intolerance towards mismatched sequences. Furthermore, it exhibits dual activation modes—responding to both DNA and RNA targets. The culmination of our research efforts has led to the development of the Hyper-Active-Verification Establishment (HAVE, 惠父). This innovation enables swift and precise human papillomavirus (HPV) diagnosis in clinical samples, providing a robust molecular diagnostic tool based on the Type I-A CRISPR-Cas3 system. Our findings contribute to understanding type I-A CRISPR-Cas3 system regulation and facilitate the creation of advanced diagnostic solutions with broad clinical applicability.

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Metadata

Authors
Hu, T., Ji, Q., Ke, X., Zhou, H., Zhang, S., Ma, S., Yu, C., Ju, W., Lu, M., Lin, Y., Ou, Y., Zhou, Y., Xiao, Y., Xu, C., & Hu, C.
Affiliations
  • Children's Hospital of Zhejiang University (CN)
  • National University of Singapore (SG)
  • Harvard University (US)
  • China Pharmaceutical University (CN)
  • Shanghai Jiao Tong University (CN)
  • ShanghaiTech University (CN)
Corresponding author
  • Zhou, Yingsi
  • Xiao, Yibei
  • Xu, Chunlong
  • Hu, Chunyi
ORCID
Venue
Communications Biology
Publisher
Springer Science and Business Media LLC
Published
2024-07-13
Volume
7
Issue
1
Pages
858-858
ISSN
2399-3642 (ISSN-L)
DOI
10.1038/s42003-024-06537-3
PMC
PMC11246428
OpenAlex
W4400606647
Cited by
24
References
2
Citations by year
2026: 8 · 2025: 14 · 2024: 2
FWCI
3.27
Type
journal article
Language
en
Open access
Licence
Locations
  • Communications Biology (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
National University of Singapore (23-0178-A0003)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Physical Sciences › Engineering › Biomedical Engineering › Biosensors and Analytical Detection
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
Keywords
autoinhibition, Cascade complex, protospacer adjacent motif, mismatch tolerance, HPV diagnosis, nucleic acid detection, molecular diagnosis
MeSH terms
Human Papillomavirus Viruses, Humans, Papillomaviridae, Papillomavirus Infections, CRISPR-Cas Systems, CRISPR-Associated Proteins

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