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1.
Nature ; 613(7944): 588-594, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36599979

RESUMEN

Bacterial abortive-infection systems limit the spread of foreign invaders by shutting down or killing infected cells before the invaders can replicate1,2. Several RNA-targeting CRISPR-Cas systems (that is, types III and VI) cause abortive-infection phenotypes by activating indiscriminate nucleases3-5. However, a CRISPR-mediated abortive mechanism that leverages indiscriminate DNase activity of an RNA-guided single-effector nuclease has yet to be observed. Here we report that RNA targeting by the type V single-effector nuclease Cas12a2 drives abortive infection through non-specific cleavage of double-stranded DNA (dsDNA). After recognizing an RNA target with an activating protospacer-flanking sequence, Cas12a2 efficiently degrades single-stranded RNA (ssRNA), single-stranded DNA (ssDNA) and dsDNA. Within cells, the activation of Cas12a2 induces an SOS DNA-damage response and impairs growth, preventing the dissemination of the invader. Finally, we harnessed the collateral activity of Cas12a2 for direct RNA detection, demonstrating that Cas12a2 can be repurposed as an RNA-guided RNA-targeting tool. These findings expand the known defensive abilities of CRISPR-Cas systems and create additional opportunities for CRISPR technologies.


Asunto(s)
Proteínas Asociadas a CRISPR , Sistemas CRISPR-Cas , ADN , ARN , Proteínas Asociadas a CRISPR/metabolismo , ADN/metabolismo , ADN de Cadena Simple/metabolismo , ARN/metabolismo , Respuesta SOS en Genética , Daño del ADN , ARN Guía de Sistemas CRISPR-Cas , Edición Génica
2.
CRISPR J ; 2(6): 434-440, 2019 12.
Artículo en Inglés | MEDLINE | ID: mdl-31809194

RESUMEN

Bacteria and archaea use CRISPR-Cas adaptive immune systems to destroy complementary nucleic acids using RNAs derived from CRISPR loci. Here, we provide the first functional evidence for type IV CRISPR-Cas, demonstrating that the system from Pseudomonas aeruginosa strain PA83 mediates RNA-guided interference against a plasmid in vivo, both clearing the plasmid and inhibiting its uptake. This interference depends on the putative NTP-dependent helicase activity of Csf4/DinG.


Asunto(s)
Sistemas CRISPR-Cas/genética , Pseudomonas aeruginosa/genética , ARN Guía de Kinetoplastida/genética , Bacterias/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , Plásmidos/genética , Interferencia de ARN/fisiología
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