Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA
| Autor: | Deborah Horst-Kreft, Joyce H.G. Lebbink, Wiggert A. van Cappellen, Dior M. J. M. Beerens, Youri Hoogstrate, Johan W. Mouton, Gert-Jan Kremers, Rob Joosten, John van der Oost, Charlie Laffeber, Duncan J. H. Gaskin, Gaurav Dugar, Rogier Louwen, Jeroen Demmers, Serena T. Bruens, Andrew P. Stubbs, Chinmoy Saha, Maarten Klunder, Peter J. van der Spek, Dik C. van Gent, Prarthana Mohanraju, Peter van Baarlen |
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| Přispěvatelé: | Medical Microbiology & Infectious Diseases, Pathology, Molecular Genetics, Radiotherapy, Biochemistry |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
DNA damage
medicine.disease_cause Campylobacter jejuni Microbiology 03 medical and health sciences chemistry.chemical_compound medicine Life Science Host-Microbe Interactomics Guide RNA VLAG 030304 developmental biology 0303 health sciences Nuclease Multidisciplinary biology 030306 microbiology Cas9 BacGen Pathogenic bacteria biology.organism_classification chemistry Streptococcus pyogenes WIAS biology.protein DNA |
| Zdroj: | Science Advances 6 (2020) 25 Science advances, 6(25):eaaz4849. American Association for the Advancement of Science Science Advances, 6(25) |
| ISSN: | 2375-2548 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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