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Cameron, Peter; Coons, Mary M; Klompe, Sanne E; Lied, Alexandra M; Smith, Stephen C; Vidal, Bastien; Donohoue, Paul D; Rotstein, Tomer; Kohrs, Bryan W; Nyer, David B; Kennedy, Rachel; Banh, Lynda M; Williams, Carolyn; Toh, Mckenzi S; Irby, Matthew J; Edwards, Leslie S; Lin, Chun-Han; Owen, Arthur L G; Künne, Tim; van der Oost, John; Brouns, Stan J J; Slorach, Euan M; Fuller, Chris K; Gradia, Scott; Kanner, Steven B; May, Andrew P; Sternberg, Samuel H
Nature biotechnology, 12/2019, Letnik: 37, Številka: 12Journal Article
Type I CRISPR-Cas systems are the most abundant adaptive immune systems in bacteria and archaea . Target interference relies on a multi-subunit, RNA-guided complex called Cascade , which recruits a trans-acting helicase-nuclease, Cas3, for target degradation . Type I systems have rarely been used for eukaryotic genome engineering applications owing to the relative difficulty of heterologous expression of the multicomponent Cascade complex. Here, we fuse Cascade to the dimerization-dependent, non-specific FokI nuclease domain and achieve RNA-guided gene editing in multiple human cell lines with high specificity and efficiencies of up to ~50%. FokI-Cascade can be reconstituted via an optimized two-component expression system encoding the CRISPR-associated (Cas) proteins on a single polycistronic vector and the guide RNA (gRNA) on a separate plasmid. Expression of the full Cascade-Cas3 complex in human cells resulted in targeted deletions of up to ~200 kb in length. Our work demonstrates that highly abundant, previously untapped type I CRISPR-Cas systems can be harnessed for genome engineering applications in eukaryotic cells.
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Dostop do baze podatkov JCR je dovoljen samo uporabnikom iz Slovenije. Vaš trenutni IP-naslov ni na seznamu dovoljenih za dostop, zato je potrebna avtentikacija z ustreznim računom AAI.
Leto | Faktor vpliva | Izdaja | Kategorija | Razvrstitev | ||||
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
Baze podatkov, v katerih je revija indeksirana
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Vir: Osebne bibliografije
in: SICRIS
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