CRISPR-CasΦ from huge phages is a hypercompact genome editor

Patrick Pausch; Basem Al-Shayeb; Ezra Bisom-Rapp; Connor A Tsuchida; Zheng Li; Brady F Cress; Gavin J Knott; Steven E Jacobsen; Jillian F Banfield; Jennifer A Doudna

doi:10.1126/science.abb1400

CRISPR-CasΦ from huge phages is a hypercompact genome editor

Science. 2020 Jul 17;369(6501):333-337. doi: 10.1126/science.abb1400.

Authors

Patrick Pausch^#^{1

2}, Basem Al-Shayeb^#^{1

3}, Ezra Bisom-Rapp⁴, Connor A Tsuchida^{1

5}, Zheng Li⁶, Brady F Cress^{1

2}, Gavin J Knott^{1

2

7}, Steven E Jacobsen^{6

8}, Jillian F Banfield^{1

9}, Jennifer A Doudna^{10

2

8

11

12

13}

Affiliations

¹ Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
² Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
³ Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, USA.
⁴ College of Natural Resources, University of California, Berkeley, Berkeley, CA, USA.
⁵ University of California, Berkeley-University of California, San Francisco Graduate Program in Bioengineering, Berkeley, CA, USA.
⁶ Department of Molecular, Cellular and Developmental Biology, University of California, Los Angeles, CA, USA.
⁷ Monash Biomedicine Discovery Institute, Department of Biochemistry and Molecular Biology, Monash University, Victoria, Australia.
⁸ Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, USA.
⁹ Department of Earth and Planetary Sciences, University of California, Berkeley, Berkeley, CA, USA.
¹⁰ Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA. doudna@berkeley.edu.
¹¹ Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA.
¹² Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
¹³ Gladstone Institute of Data Science and Biotechnology, Gladstone Institutes, San Francisco, CA, USA.

^# Contributed equally.

Abstract

CRISPR-Cas systems are found widely in prokaryotes, where they provide adaptive immunity against virus infection and plasmid transformation. We describe a minimal functional CRISPR-Cas system, comprising a single ~70-kilodalton protein, CasΦ, and a CRISPR array, encoded exclusively in the genomes of huge bacteriophages. CasΦ uses a single active site for both CRISPR RNA (crRNA) processing and crRNA-guided DNA cutting to target foreign nucleic acids. This hypercompact system is active in vitro and in human and plant cells with expanded target recognition capabilities relative to other CRISPR-Cas proteins. Useful for genome editing and DNA detection but with a molecular weight half that of Cas9 and Cas12a genome-editing enzymes, CasΦ offers advantages for cellular delivery that expand the genome editing toolbox.

CRISPR-CasΦ from huge phages is a hypercompact genome editor

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