Transposon-encoded CRISPR–Cas systems direct RNA-guided DNA integration
| Autor: | Sanne E. Klompe, Phuc Leo H. Vo, Samuel H. Sternberg, Tyler S. Halpin-Healy |
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| Rok vydání: | 2019 |
| Předmět: |
Transposable element
0303 health sciences Multidisciplinary Computational biology Biology Genome DNA sequencing 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine chemistry CRISPR DNA Integration Guide RNA Mobile genetic elements 030217 neurology & neurosurgery DNA 030304 developmental biology |
| Zdroj: | Nature. 571:219-225 |
| ISSN: | 1476-4687 0028-0836 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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