CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity

Author:

Chen Janice S.1ORCID,Ma Enbo1,Harrington Lucas B.1,Da Costa Maria2ORCID,Tian Xinran3ORCID,Palefsky Joel M.2ORCID,Doudna Jennifer A.13456ORCID

Affiliation:

1. Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.

2. Department of Medicine, University of California, San Francisco, San Francisco, CA 94143, USA.

3. Department of Chemistry, University of California, Berkeley, Berkeley, CA 94720, USA.

4. Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA 94704, USA.

5. Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA 94720, USA.

6. Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.

Abstract

Taking CRISPR technology further CRISPR techniques are allowing the development of technologies for nucleic acid detection (see the Perspective by Chertow). Taking advantages of the distinctive enzymatic properties of CRISPR enzymes, Gootenberg et al. developed an improved nucleic acid detection technology for multiplexed quantitative and highly sensitive detection, combined with lateral flow for visual readout. Myhrvold et al. added a sample preparation protocol to create a field-deployable viral diagnostic platform for rapid detection of specific strains of pathogens in clinical samples. Cas12a (also known as Cpf1), a type V CRISPR protein, cleaves double-stranded DNA and has been adapted for genome editing. Chen et al. discovered that Cas12a also processes single-stranded DNA threading activity. A technology platform based on this activity detected human papillomavirus in patient samples with high sensitivity. Science , this issue p. 439 , p. 444 , p. 436 ; see also p. 381

Funder

National Science Foundation

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

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