Print Email Facebook Twitter Cas4 Facilitates PAM-Compatible Spacer Selection during CRISPR Adaptation Title Cas4 Facilitates PAM-Compatible Spacer Selection during CRISPR Adaptation Author Kieper, S.N. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Almendros Romero, C. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Behler, Juliane (Albert-Ludwigs-Universität Freiburg) McKenzie, R. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Luzia de Nobrega, F. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) van Eijkeren-Haagsma, A.C. (TU Delft BN/Technici en Analisten; Kavli institute of nanoscience Delft) Vink, J.N.A. (TU Delft BN/Stan Brouns Lab; Kavli institute of nanoscience Delft) Hess, Wolfgang R. (Albert-Ludwigs-Universität Freiburg) Brouns, S.J.J. (TU Delft BN/Stan Brouns Lab; Wageningen University & Research; Kavli institute of nanoscience Delft) Date 2018 Abstract CRISPR-Cas systems adapt their immunological memory against their invaders by integrating short DNA fragments into clustered regularly interspaced short palindromic repeat (CRISPR) loci. While Cas1 and Cas2 make up the core machinery of the CRISPR integration process, various class I and II CRISPR-Cas systems encode Cas4 proteins for which the role is unknown. Here, we introduced the CRISPR adaptation genes cas1, cas2, and cas4 from the type I-D CRISPR-Cas system of Synechocystis sp. 6803 into Escherichia coli and observed that cas4 is strictly required for the selection of targets with protospacer adjacent motifs (PAMs) conferring I-D CRISPR interference in the native host Synechocystis. We propose a model in which Cas4 assists the CRISPR adaptation complex Cas1-2 by providing DNA substrates tailored for the correct PAM. Introducing functional spacers that target DNA sequences with the correct PAM is key to successful CRISPR interference, providing a better chance of surviving infection by mobile genetic elements. Kieper et al. demonstrate that the ubiquitous protein Cas4 assists Cas1 and Cas2 in the selection of new CRISPR spacers with a PAM licensing efficient CRISPR interference. Subject Cas4CRISPR adaptationspacer acquisitiontype I-D CRISPR-Cas system To reference this document use: http://resolver.tudelft.nl/uuid:2daac99d-c3d9-4874-a3a8-2490ba05834d DOI https://doi.org/10.1016/j.celrep.2018.02.103 ISSN 2211-1247 Source Cell Reports, 22 (13), 3377-3384 Part of collection Institutional Repository Document type journal article Rights © 2018 S.N. Kieper, C. Almendros Romero, Juliane Behler, R. McKenzie, F. Luzia de Nobrega, A.C. van Eijkeren-Haagsma, J.N.A. Vink, Wolfgang R. Hess, S.J.J. Brouns Files PDF 1_s2.0_S2211124718303139_main.pdf 1.18 MB Close viewer /islandora/object/uuid:2daac99d-c3d9-4874-a3a8-2490ba05834d/datastream/OBJ/view