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CRISPR pp 147-159 | Cite as

Rapid Multiplex Creation of Escherichia coli Strains Capable of Interfering with Phage Infection Through CRISPR

  • Alexandra Strotksaya
  • Ekaterina Semenova
  • Ekaterina Savitskaya
  • Konstantin Severinov
Part of the Methods in Molecular Biology book series (MIMB, volume 1311)

Abstract

In Escherichia coli, acquisition of new spacers in the course of CRISPR-Cas adaptation is dramatically stimulated by preexisting partial matches between a bacterial CRISPR cassette spacer and a protospacer sequence in the DNA of the infecting bacteriophage or plasmid. This phenomenon, which we refer to as “priming,” can be used for very simple and rapid construction of multiple E. coli strains capable of targeting, through CRISPR interference, any phage or plasmid of interest. Availability of such strains should allow rapid progress in the analysis of CRISPR-Cas system function against diverse mobile genetic elements.

Key words

Escherichia coli CRISPR Cas proteins Bacteriophage Spacers Strain construction 

Notes

Acknowledgments

This work was supported by National Institutes of Health grant R01 GM10407, Molecular and Cell Biology Program grant from the Russian Academy of Sciences Presidium, and Ministry of Education and Science of Russian Federation project 14.B25.31.0004 and Russian Science Foundation grant 14-14-00988 (to KS) and Russian Foundation for Basic Research grant 14-04-00916 (to Ekaterina Savitskaya).

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Alexandra Strotksaya
    • 1
    • 2
    • 4
  • Ekaterina Semenova
    • 3
  • Ekaterina Savitskaya
    • 1
    • 4
  • Konstantin Severinov
    • 1
    • 2
    • 3
    • 5
  1. 1.Skolkovo Institute of Science and TechnologyMoscowRussia
  2. 2.St. Petersburg State Polytechnical UniversitySt. PetersburgRussia
  3. 3.Waksman Institute of MicrobiologyPiscatawayUSA
  4. 4.Institute of Molecular GeneticsRussian Academy of SciencesMoscowRussia
  5. 5.Institute of Gene BiologyRussian Academy of SciencesMoscowRussia

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