Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA.
Sci Adv
; 6(25): eaaz4849, 2020 06.
Article
en En
| MEDLINE
| ID: mdl-32596446
ABSTRACT
CRISPR-Cas9 systems are enriched in human pathogenic bacteria and have been linked to cytotoxicity by an unknown mechanism. Here, we show that upon infection of human cells, Campylobacter jejuni secretes its Cas9 (CjeCas9) nuclease into their cytoplasm. Next, a native nuclear localization signal enables CjeCas9 nuclear entry, where it catalyzes metal-dependent nonspecific DNA cleavage leading to cell death. Compared to CjeCas9, native Cas9 of Streptococcus pyogenes (SpyCas9) is more suitable for guide-dependent editing. However, in human cells, native SpyCas9 may still cause some DNA damage, most likely because of its ssDNA cleavage activity. This side effect can be completely prevented by saturation of SpyCas9 with an appropriate guide RNA, which is only partially effective for CjeCas9. We conclude that CjeCas9 plays an active role in attacking human cells rather than in viral defense. Moreover, these unique catalytic features may therefore make CjeCas9 less suitable for genome editing applications.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Campylobacter jejuni
/
Proteína 9 Asociada a CRISPR
Tipo de estudio:
Etiology_studies
Límite:
Humans
Idioma:
En
Año:
2020
Tipo del documento:
Article