期刊
NUCLEIC ACIDS RESEARCH
卷 42, 期 9, 页码 5907-5916出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gku226
关键词
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资金
- National Institutes of Health (NIH) [GM10407]
- Russian Foundation for Basic Research Foundation [11-04-01373-a, 14-04-00916]
- Ministry of Education and Science of Russian Federation [14.B25.31.0004]
- Ministry of Education and Science of Russion Federation [14.B25.31.0004]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM104071] Funding Source: NIH RePORTER
During the process of prokaryotic CRISPR adaptation, a copy of a segment of foreign deoxyribonucleic acid referred to as protospacer is added to the CRISPR cassette and becomes a spacer. When a protospacer contains a neighboring target interference motif, the specific small CRISPR ribonucleic acid (crRNA) transcribed from expanded CRISPR cassette can protect a prokaryotic cell from virus infection or plasmid transformation and conjugation. We show that in Escherichia coli, a vast majority of plasmid protospacers generate spacers integrated in CRISPR cassette in two opposing orientations, leading to frequent appearance of complementary spacer pairs in a population of cells that underwent CRISPR adaptation. When a protospacer contains a spacer acquisition motif AAG, spacer orientation that generates functional protective crRNA is strongly preferred. All other protospacers give rise to spacers oriented in both ways at comparable frequencies. This phenomenon increases the repertoire of available spacers and should make it more likely that a protective crRNA is formed as a result of CRISPR adaptation.
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