TY - JOUR
T1 - Rapid multiplex creation of Escherichia coli strains capable of interfering with phage infection through CRISPR
AU - Strotksaya, Alexandra
AU - Semenova, Ekaterina
AU - Savitskaya, Ekaterina
AU - Severinov, Konstantin
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2015.
PY - 2015
Y1 - 2015
N2 - 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.
AB - 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.
KW - Bacteriophage
KW - CRISPR
KW - Cas proteins
KW - Escherichia coli
KW - Spacers
KW - Strain construction
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U2 - 10.1007/978-1-4939-2687-9_9
DO - 10.1007/978-1-4939-2687-9_9
M3 - Article
C2 - 25981471
AN - SCOPUS:85025147704
SN - 1064-3745
VL - 1311
SP - 147
EP - 159
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -