The DOI matches its Crossref record; 68 of the 68 references it cites were found in a registry or an index.
Photocontrolled crRNA activation enables robust CRISPR-Cas12a diagnostics
Open accessAbstract
CRISPR diagnostics based on nucleic acid amplification faces barriers to its commercial use, such as contamination risks and insufficient sensitivity. Here, we propose a robust solution involving optochemical control of CRISPR RNA (crRNA) activation in CRISPR detection. Based on this strategy, recombinase polymerase amplification (RPA) and CRISPR-Cas12a detection systems can be integrated into a completely closed test tube. crRNA can be designed to be temporarily inactivated so that RPA is not affected by Cas12a cleavage. After the RPA reaction is completed, the CRISPR-Cas12a detection system is activated under rapid light irradiation. This photocontrolled, fully closed CRISPR diagnostic system avoids contamination risks and exhibits a more than two orders of magnitude improvement in sensitivity compared with the conventional one-pot assay. This photocontrolled CRISPR method was applied to the clinical detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA, achieving detection sensitivity and specificity comparable to those of PCR. Furthermore, a compact and automatic photocontrolled CRISPR detection device was constructed.
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Cites
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Metadata
- Authors
- Hu, M., Qiu, Z., Bi, Z., Tian, T., Jiang, Y., & Zhou, X.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2022-06-21
- Volume
- 119
- Issue
- 26
- Pages
- e2202034119-e2202034119
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W4283219719
- Cited by
- 293
- References
- 68
- FWCI
- 13.90
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Hu, M., Qiu, Z., Bi, Z., Tian, T., Jiang, Y., & Zhou, X. (2022). Photocontrolled crRNA activation enables robust CRISPR-Cas12a diagnostics. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.2202034119
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