Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 509410
Title Cas3-Derived Target DNA Degradation Fragments Fuel Primed CRISPR Adaptation
Author(s) Künne, Tim; Kieper, Sebastian N.; Bannenberg, Jasper W.; Vogel, Anne; Miellet, Willem R.; Klein, Misha; Depken, Martin; Suarez-Diez, Maria; Brouns, Stan J.J.
Source Molecular Cell 63 (2016)5. - ISSN 1097-2765 - p. 852 - 864.
DOI http://dx.doi.org/10.1016/j.molcel.2016.07.011
Department(s) Microbiological Laboratory
Systems and Synthetic Biology
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2016
Keyword(s) adaptive immunity - Cas1 - Cas2 - Cas3 - Cascade - CRISPR-Cas - interference - phage resistance - priming - spacer acquisition
Abstract

Prokaryotes use a mechanism called priming to update their CRISPR immunological memory to rapidly counter revisiting, mutated viruses, and plasmids. Here we have determined how new spacers are produced and selected for integration into the CRISPR array during priming. We show that Cas3 couples CRISPR interference to adaptation by producing DNA breakdown products that fuel the spacer integration process in a two-step, PAM-associated manner. The helicase-nuclease Cas3 pre-processes target DNA into fragments of about 30–100 nt enriched for thymine-stretches in their 3′ ends. The Cas1-2 complex further processes these fragments and integrates them sequence-specifically into CRISPR repeats by coupling of a 3′ cytosine of the fragment. Our results highlight that the selection of PAM-compliant spacers during priming is enhanced by the combined sequence specificities of Cas3 and the Cas1-2 complex, leading to an increased propensity of integrating functional CTT-containing spacers.

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