Rxivist logo

Rxivist combines preprints from bioRxiv with data from Twitter to help you find the papers being discussed in your field. Currently indexing 73,960 bioRxiv papers from 321,862 authors.

Selective Prespacer Processing Ensures Precise CRISPR-Cas Adaptation

By Sungchul Kim, Luuk Loeff, Sabina Colombo, Stan J.J. Brouns, Chirlmin Joo

Posted 16 Apr 2019
bioRxiv DOI: 10.1101/608976

CRISPR-Cas immunity protects prokaryotes against foreign genetic elements. CRISPR-Cas uses the highly conserved Cas1-Cas2 complex to establish inheritable memory (spacers). It remains elusive how Cas1-Cas2 acquires spacers from cellular DNA fragments (prespacers) and how it integrates them into the CRISPR array in the correct orientation. By using the high spatiotemporal resolution of single-molecule fluorescence, we reveal that Cas1-Cas2 obtains prespacers in various forms including single-stranded DNA and partial duplexes by selecting them in the DNA-length and PAM-dependent manner. Furthermore, we identify DnaQ exonucleases as enzymes that can mature the Cas1- Cas2-loaded precursor prespacers into an integration-competent size. Cas1-Cas2 protects the PAM sequence from maturation, which results in the production of asymmetrically trimmed prespacers and subsequent spacer integration in the correct orientation. This kinetic coordination in prespacer selection and PAM trimming provides comprehensive understanding of the mechanisms that underlie the integration of functional spacers in the CRISPR array.

Download data

  • Downloaded 806 times
  • Download rankings, all-time:
    • Site-wide: 12,340 out of 73,960
    • In biophysics: 391 out of 3,135
  • Year to date:
    • Site-wide: 13,120 out of 73,960
  • Since beginning of last month:
    • Site-wide: 13,120 out of 73,960

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

Sign up for the Rxivist weekly newsletter! (Click here for more details.)


News