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Nature. 2019 Jul;571(7764):219-225. doi: 10.1038/s41586-019-1323-z. Epub 2019 Jun 12.

Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA integration.

Author information

1
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA.
2
Department of Pharmacology, Columbia University, New York, NY, USA.
3
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, NY, USA. shsternberg@gmail.com.

Abstract

Conventional CRISPR-Cas systems maintain genomic integrity by leveraging guide RNAs for the nuclease-dependent degradation of mobile genetic elements, including plasmids and viruses. Here we describe a notable inversion of this paradigm, in which bacterial Tn7-like transposons have co-opted nuclease-deficient CRISPR-Cas systems to catalyse RNA-guided integration of mobile genetic elements into the genome. Programmable transposition of Vibrio cholerae Tn6677 in Escherichia coli requires CRISPR- and transposon-associated molecular machineries, including a co-complex between the DNA-targeting complex Cascade and the transposition protein TniQ. Integration of donor DNA occurs in one of two possible orientations at a fixed distance downstream of target DNA sequences, and can accommodate variable length genetic payloads. Deep-sequencing experiments reveal highly specific, genome-wide DNA insertion across dozens of unique target sites. This discovery of a fully programmable, RNA-guided integrase lays the foundation for genomic manipulations that obviate the requirements for double-strand breaks and homology-directed repair.

PMID:
31189177
DOI:
10.1038/s41586-019-1323-z

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