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

Sanne E. Klompe, Phuc L. H. Vo, Tyler S. Halpin-Healy and Samuel H. Sternberg ()
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Sanne E. Klompe: Columbia University
Phuc L. H. Vo: Columbia University
Tyler S. Halpin-Healy: Columbia University
Samuel H. Sternberg: Columbia University

Nature, 2019, vol. 571, issue 7764, 219-225

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41586-019-1323-z

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