The DOI matches its Crossref record; 48 of the 48 references it cites were found in a registry or an index.
CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity
Open accessPublished version of a preprint
Abstract
CRISPR-Cas12a (Cpf1) proteins are RNA-guided enzymes that bind and cut DNA as components of bacterial adaptive immune systems. Like CRISPR-Cas9, Cas12a has been harnessed for genome editing on the basis of its ability to generate targeted, double-stranded DNA breaks. Here we show that RNA-guided DNA binding unleashes indiscriminate single-stranded DNA (ssDNA) cleavage activity by Cas12a that completely degrades ssDNA molecules. We find that target-activated, nonspecific single-stranded deoxyribonuclease (ssDNase) cleavage is also a property of other type V CRISPR-Cas12 enzymes. By combining Cas12a ssDNase activation with isothermal amplification, we create a method termed DNA endonuclease-targeted CRISPR trans reporter (DETECTR), which achieves attomolar sensitivity for DNA detection. DETECTR enables rapid and specific detection of human papillomavirus in patient samples, thereby providing a simple platform for molecular diagnostics.
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“Cas12a enzymes are single crRNA-guided endonucleases measuring 3.9 kb”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [78]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“their distinguishable differences from Cas9 include: a T nucleotide-rich PAM site (5′-TTTV), self-catalysis of crRNA maturation, and a staggered dsDNA cut [76,77,78,79].”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [78]
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“Additionally, Cas12a′s trans-nuclease activity is utilized to detect sequence-specific nucleic acid (Chen et al., 2018).”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Chen et al., 2018
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In the citing paper
“their distinguishable differences from Cas9 include: a T nucleotide-rich PAM site (5′-TTTV), self-catalysis of crRNA maturation, and a staggered dsDNA cut [76,77,78,79].”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [78]
The cited source says what the claim attributes to it.
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“their distinguishable differences from Cas9 include: a T nucleotide-rich PAM site (5′-TTTV), self-catalysis of crRNA maturation, and a staggered dsDNA cut [76,77,78,79].”
From Viral Delivery Systems for CRISPR (2019) · cites this paper as [78]
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Citations
Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116
Self-citationThe new frontier of genome engineering with CRISPR-Cas9
Doudna, J. A., & Charpentier, E. (2014) · Science · doi:10.1126/science.1258096 · Cited by 7,062
Self-citationCRISPR Provides Acquired Resistance Against Viruses in Prokaryotes
Barrangou, R., Fremaux, C., Deveau, H., Richards, M., Boyaval, P., Moineau, S., Romero, D. A., & Horvath, P. (2007) · Science · doi:10.1126/science.1138140 · Cited by 5,947
Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., Volz, S. E., Joung, J., van der Oost, J., Regev, A., et al. (2015) · Cell · doi:10.1016/j.cell.2015.09.038 · Cited by 4,761
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. K., Donghia, N. M., Daringer, N. M., Freije, C. A., et al. (2017) · Science · doi:10.1126/science.aam9321 · Cited by 3,887
An updated evolutionary classification of CRISPR–Cas systems
Makarova, K. S., Wolf, Y. I., Alkhnbashi, O. S., Costa, F., Shah, S. A., Saunders, S. J., Barrangou, R., Brouns, S. J. J., Charpentier, E., Haft, D. H., et al. (2015) · Nature Reviews Microbiology · doi:10.1038/nrmicro3569 · Cited by 2,480
C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector
Abudayyeh, O. O., Gootenberg, J. S., Konermann, S., Joung, J., Slaymaker, I. M., Cox, D. B. T., Shmakov, S., Makarova, K. S., Semenova, E., Minakhin, L., et al. (2016) · Science · doi:10.1126/science.aaf5573 · Cited by 2,336
Crystal structure of Cas9 in complex with guide RNA and target DNA
Nishimasu, H., Ran, F. A., Hsu, P. D., Konermann, S., Shehata, S. I., Dohmae, N., Ishitani, R., Zhang, F., & Nureki, O. (2014) · Cell · doi:10.1016/j.cell.2014.02.001 · Cited by 2,306
DNA interrogation by the CRISPR RNA-guided endonuclease Cas9
Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C., & Doudna, J. A. (2014) · Nature · doi:10.1038/nature13011 · Cited by 1,822
Self-citationCRISPR Interference Limits Horizontal Gene Transfer in Staphylococci by Targeting DNA
Marraffini, L. A., & Sontheimer, E. J. (2008) · Science · doi:10.1126/science.1165771 · Cited by 1,642
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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. C., Sotomayor-González, A., et al. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0513-4 · Cited by 2,777
Self-citationMultiplexed 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 2,679
Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors
Anzalone, A. V., Koblan, L. W., & Liu, D. R. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0561-9 · Cited by 2,237
Li, H., Yang, Y., Hong, W., Huang, M., Wu, M., & Zhao, X. (2020) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-019-0089-y · Cited by 1,829
SHERLOCK: nucleic acid detection with CRISPR nucleases
Kellner, M. J., Koob, J. G., Gootenberg, J. S., Abudayyeh, O. O., & Zhang, F. (2019) · Nature Protocols · doi:10.1038/s41596-019-0210-2 · Cited by 1,699
Metadata
- Authors
- Chen, J. S., Ma, E., Harrington, L. B., Da Costa, M., Tian, X., Palefsky, J. M., & Doudna, J. A.
- Affiliations
- University of California, Berkeley (US)
- Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.
- University of California, San Francisco (US)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Chen, Janice S.
- Ma, Enbo
- Harrington, Lucas B.
- Doudna, Jennifer A.
- ORCID
- Venue
- Science
- Published
- 2018-04-27
- Volume
- 360
- Issue
- 6387
- Pages
- 436-439
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- PMC
- PMC6628903
- OpenAlex
- W2787024379
- Cited by
- 4,507
- References
- 48
- Citations by year
- 2026: 759 · 2025: 816 · 2024: 736 · 2023: 643 · 2022: 624 · 2021: 452 · 2020: 271 · 2019: 135 · 2018: 67
- FWCI
- 82.27
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Locations
- Science (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- National Science Foundation (MCB-1244557)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- CRISPR/Cas12a, single-stranded DNA cleavage, isothermal amplification, DETECTR, DNA detection, HPV testing, molecular diagnosis, genome editing
- MeSH terms
- Clostridiales, Eubacteriales, Bacterial Proteins, DNA, Single-Stranded, Endodeoxyribonucleases, Endonucleases, Kinetics, Substrate Specificity, DNA Cleavage, CRISPR-Cas Systems, CRISPR-Associated Proteins
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