Abstract：
:Type I CRISPR-Cas loci provide prokaryotes with a nucleic-acid-based adaptive immunity against foreign DNA. Immunity involves adaptation, the integration of ~30-bp DNA fragments, termed prespacers, into the CRISPR array as spacers, and interference, the targeted degradation of DNA containing a protospacer. Interference-driven DNA degradation can be coupled with primed adaptation, in which spacers are acquired from DNA surrounding the targeted protospacer. Here we develop a method for strand-specific, high-throughput sequencing of DNA fragments, FragSeq, and apply this method to identify DNA fragments accumulated in Escherichia coli cells undergoing robust primed adaptation by a type I-E or type I-F CRISPR-Cas system. The detected fragments have sequences matching spacers acquired during primed adaptation and function as spacer precursors when introduced exogenously into cells by transformation. The identified prespacers contain a characteristic asymmetrical structure that we propose is a key determinant of integration into the CRISPR array in an orientation that confers immunity.
journal_name
Nat Communjournal_title
Nature communicationsauthors
Shiriaeva AA,Savitskaya E,Datsenko KA,Vvedenskaya IO,Fedorova I,Morozova N,Metlitskaya A,Sabantsev A,Nickels BE,Severinov K,Semenova Edoi
10.1038/s41467-019-12417-wsubject
Has Abstractpub_date
2019-10-10 00:00:00pages
4603issue
1issn
2041-1723pii
10.1038/s41467-019-12417-wjournal_volume
10pub_type
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