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Musharova O, Sitnik V, Vlot M, et al. Systematic analysis of Type I-E Escherichia coli CRISPR-Cas PAM sequences ability to promote interference and primed adaptation. Mol Microbiol. 2019;111(6):1558-1570doi: 10.1111/mmi.14237.
Musharova, O., Sitnik, V., Vlot, M., Savitskaya, E., Datsenko, K. A., Krivoy, A., Fedorov, I., Semenova, E., Brouns, S. J. J., & Severinov, K. (2019). Systematic analysis of Type I-E Escherichia coli CRISPR-Cas PAM sequences ability to promote interference and primed adaptation. Molecular microbiology, 111(6), 1558-1570. https://doi.org/10.1111/mmi.14237
Musharova, Olga, et al. "Systematic analysis of Type I-E Escherichia coli CRISPR-Cas PAM sequences ability to promote interference and primed adaptation." Molecular microbiology vol. 111,6 (2019): 1558-1570. doi: https://doi.org/10.1111/mmi.14237
Musharova O, Sitnik V, Vlot M, Savitskaya E, Datsenko KA, Krivoy A, Fedorov I, Semenova E, Brouns SJJ, Severinov K. Systematic analysis of Type I-E Escherichia coli CRISPR-Cas PAM sequences ability to promote interference and primed adaptation. Mol Microbiol. 2019 Jun;111(6):1558-1570. doi: 10.1111/mmi.14237. Epub 2019 Apr 06. PMID: 30875129; PMCID: PMC6568314.
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Mol Microbiol. 2019 Jun;111(6):1558-1570. doi: 10.1111/mmi.14237. Epub 2019 Apr 06.

Systematic analysis of Type I-E Escherichia coli CRISPR-Cas PAM sequences ability to promote interference and primed adaptation.

Molecular microbiology

Olga Musharova, Vasily Sitnik, Marnix Vlot, Ekaterina Savitskaya, Kirill A Datsenko, Andrey Krivoy, Ivan Fedorov, Ekaterina Semenova, Stan J J Brouns, Konstantin Severinov

Affiliations

  1. Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow, 121205, Russia.
  2. Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, 123182, Russia.
  3. Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Stippeneng 4, Wageningen, 6708 WE, The Netherlands.
  4. Department of Biological Sciences, Purdue University, West Lafayette, IN, 47907, USA.
  5. Waksman Institute, Rutgers, The State University of New Jersey, Piscataway, NJ, 08854, USA.
  6. Department of Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, Delft, 2629 HZ, The Netherlands.

PMID: 30875129 PMCID: PMC6568314 DOI: 10.1111/mmi.14237

Abstract

CRISPR interference occurs when a protospacer recognized by the CRISPR RNA is destroyed by Cas effectors. In Type I CRISPR-Cas systems, protospacer recognition can lead to «primed adaptation» - acquisition of new spacers from in cis located sequences. Type I CRISPR-Cas systems require the presence of a trinucleotide protospacer adjacent motif (PAM) for efficient interference. Here, we investigated the ability of each of 64 possible trinucleotides located at the PAM position to induce CRISPR interference and primed adaptation by the Escherichia coli Type I-E CRISPR-Cas system. We observed clear separation of PAM variants into three groups: those unable to cause interference, those that support rapid interference and those that lead to reduced interference that occurs over extended periods of time. PAM variants unable to support interference also did not support primed adaptation; those that supported rapid interference led to no or low levels of adaptation, while those that caused attenuated levels of interference consistently led to highest levels of adaptation. The results suggest that primed adaptation is fueled by the products of CRISPR interference. Extended over time interference with targets containing «attenuated» PAM variants provides a continuous source of new spacers leading to high overall level of spacer acquisition.

© 2019 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd.

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