The DOI matches its Crossref record; 39 of the 39 references it cites were found in a registry or an index.
Accelerated RNA detection using tandem CRISPR nucleases
Open accessAbstract
Direct, amplification-free detection of RNA has the potential to transform molecular diagnostics by enabling simple on-site analysis of human or environmental samples. CRISPR–Cas nucleases offer programmable RNA-guided RNA recognition that triggers cleavage and release of a fluorescent reporter molecule, but long reaction times hamper their detection sensitivity and speed. Here, we show that unrelated CRISPR nucleases can be deployed in tandem to provide both direct RNA sensing and rapid signal generation, thus enabling robust detection of ~30 molecules per µl of RNA in 20 min. Combining RNA-guided Cas13 and Csm6 with a chemically stabilized activator creates a one-step assay that can detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA extracted from respiratory swab samples with quantitative reverse transcriptase PCR (qRT–PCR)-derived cycle threshold (Ct) values up to 33, using a compact detector. This Fast Integrated Nuclease Detection In Tandem (FIND-IT) approach enables sensitive, direct RNA detection in a format that is amenable to point-of-care infection diagnosis as well as to a wide range of other diagnostic or research applications. Deploying two unrelated CRISPR nucleases in tandem, with multiplexed CRISPR RNAs and a chemically stabilized activator, creates a simple, one-step assay that can rapidly detect attomolar concentrations of RNA without needing target amplification.
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Cites
Nucleic acid detection with CRISPR-Cas13a/C2c2
Gootenberg, J. S., Abudayyeh, O. O., Lee, J. W., Essletzbichler, P., Dy, A. J., Joung, J., Verdine, V., Donghia, N., Daringer, N. M., Freije, C. A., et al. (2017) · Science · doi:10.1126/science.aam9321 · Cited by 3,844
CRISPR–Cas12-based detection of SARS-CoV-2
Broughton, J. P., Deng, X., Yu, G., Fasching, C. L., Servellita, V., Singh, J., Miao, X., Streithorst, J. A., Granados, A., Sotomayor-Gonzalez, A., et al. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0513-4 · Cited by 2,717
Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6
Gootenberg, J. S., Abudayyeh, O. O., Kellner, M. J., Joung, J., Collins, J. J., & Zhang, F. (2018) · Science · doi:10.1126/science.aaq0179 · Cited by 2,638
DNA Detection Using Recombination Proteins
Piepenburg, O., Williams, C. H., Stemple, D. L., & Armes, N. A. (2006) · PLoS Biology · doi:10.1371/journal.pbio.0040204 · Cited by 2,482
Self-citationEvolutionary classification of CRISPR–Cas systems: a burst of class 2 and derived variants
Makarova, K. S., Wolf, Y. I., Iranzo, J., Shmakov, S. A., Alkhnbashi, O. S., Brouns, S. J. J., Charpentier, E., Cheng, D., Haft, D. H., Horvath, P., et al. (2020) · Nature Reviews Microbiology · doi:10.1038/s41579-019-0299-x · Cited by 2,445
Accelerated reaction by loop-mediated isothermal amplification using loop primers
Nagamine, K., Hase, T., & Notomi, T. (2002) · Molecular and Cellular Probes · doi:10.1006/mcpr.2002.0415 · Cited by 2,045
Khalifa, A. A. Z., Ichikawa, M., Dai, D., Kubo, S., Black, C. S., Peri, K., McAlear, T. S., Veyron, S., Yang, S. K., Vargas, J., et al. (2020) · eLife · doi:10.7554/elife.52760 · Cited by 1,676
CRISPR-Cas guides the future of genetic engineering
Knott, G. J., & Doudna, J. A. (2018) · Science · doi:10.1126/science.aat5011 · Cited by 1,410
Self-citationTwo distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection
East-Seletsky, A., O’Connell, M. R., Knight, S. C., Burstein, D., Cate, J. H. D., Tjian, R., & Doudna, J. A. (2016) · Nature · doi:10.1038/nature19802 · Cited by 1,236
Self-citationTest sensitivity is secondary to frequency and turnaround time for COVID-19 screening
Larremore, D. B., Wilder, B., Lester, E., Shehata, S., Burke, J. M., Hay, J. A., Tambe, M., Mina, M. J., & Parker, R. (2021) · Science Advances · doi:10.1126/sciadv.abd5393 · Cited by 1,216
Cited by
(2023) · doi:10.1016/j.xcrm.2023.101037 · Cited by 75
Metadata
- Authors
- Liu, T. Y., Knott, G. J., Smock, D. C. J., Desmarais, J. J., Son, S., Bhuiya, A., Jakhanwal, S., Prywes, N., Agrawal, S., Díaz de León Derby, M., Switz, N. A., Armstrong, M., Harris, A. R., Charles, E. J., Thornton, B. W., Fozouni, P., Shu, J., Stephens, S. I., Kumar, G. R., Zhao, C., Mok, A., Iavarone, A. T., Escajeda, A. M., McIntosh, R., Kim, S., Dugan, E. J., IGI Testing Consortium, Hamilton, J. R., Lin-Shiao, E., Stahl, E. C., Tsuchida, C. A., Moehle, E. A., Giannikopoulos, P., McElroy, M., McDevitt, S., Zur, A., Sylvain, I., Ciling, A., Zhu, M., Williams, C., Baldwin, A., Pollard, K. S., Tan, M. X., Ott, M., Fletcher, D. A., Lareau, L. F., Hsu, P. D., Savage, D. F., & Doudna, J. A.
- Venue
- Nature Chemical Biology
- Published
- 2021-08-05
- Volume
- 17
- Issue
- 9
- Pages
- 982-988
- ISSN
- 1552-4450 (ISSN-L)
- 1552-4469
- OpenAlex
- W3188079964
- Cited by
- 268
- References
- 39
- FWCI
- 11.54
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Liu, T. Y., Knott, G. J., Smock, D. C. J., Desmarais, J. J., Son, S., Bhuiya, A., Jakhanwal, S., Prywes, N., Agrawal, S., Díaz de León Derby, M., Switz, N. A., Armstrong, M., Harris, A. R., Charles, E. J., Thornton, B. W., Fozouni, P., Shu, J., Stephens, S. I., Kumar, G. R., . . . Doudna, J. A. (2021). Accelerated RNA detection using tandem CRISPR nucleases. Nature Chemical Biology. https://doi.org/10.1038/s41589-021-00842-2
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