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Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA.
Sci Adv. 2020 Jun; 6(25):eaaz4849.SA

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.

Authors+Show Affiliations

Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands.Clinical Bioinformatics, Department of Pathology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Institute of Molecular Infection Biology (IMIB)/Research Center for Infectious Diseases (ZINF), University of Würzburg, Würzburg, Germany.Clinical Bioinformatics, Department of Pathology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Optical Imaging Center, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Optical Imaging Center, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Department of Molecular Genetics, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Department of Molecular Genetics, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands. Department of Radiation Oncology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Department of Molecular Genetics, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Institute of Food Research, Gut Health and Food Safety Programme, Norwich Research Park, Norwich, UK.Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands.Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands.Proteomics Center, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Department of Molecular Genetics, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Clinical Bioinformatics, Department of Pathology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.Laboratory of Microbiology, Wageningen University, Wageningen, Netherlands.Host-Microbe Interactomics Group, University of Wageningen, Wageningen, Netherlands.Department of Medical Microbiology and Infectious Diseases, Erasmus MC, University Medical Center Rotterdam, Rotterdam, Netherlands.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32596446

Citation

Saha, Chinmoy, et al. "Guide-free Cas9 From Pathogenic Campylobacter Jejuni Bacteria Causes Severe Damage to DNA." Science Advances, vol. 6, no. 25, 2020, pp. eaaz4849.
Saha C, Mohanraju P, Stubbs A, et al. Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA. Sci Adv. 2020;6(25):eaaz4849.
Saha, C., Mohanraju, P., Stubbs, A., Dugar, G., Hoogstrate, Y., Kremers, G. J., van Cappellen, W. A., Horst-Kreft, D., Laffeber, C., Lebbink, J. H. G., Bruens, S., Gaskin, D., Beerens, D., Klunder, M., Joosten, R., Demmers, J. A. A., van Gent, D., Mouton, J. W., van der Spek, P. J., ... Louwen, R. (2020). Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA. Science Advances, 6(25), eaaz4849. https://doi.org/10.1126/sciadv.aaz4849
Saha C, et al. Guide-free Cas9 From Pathogenic Campylobacter Jejuni Bacteria Causes Severe Damage to DNA. Sci Adv. 2020;6(25):eaaz4849. PubMed PMID: 32596446.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Guide-free Cas9 from pathogenic Campylobacter jejuni bacteria causes severe damage to DNA. AU - Saha,Chinmoy, AU - Mohanraju,Prarthana, AU - Stubbs,Andrew, AU - Dugar,Gaurav, AU - Hoogstrate,Youri, AU - Kremers,Gert-Jan, AU - van Cappellen,Wiggert A, AU - Horst-Kreft,Deborah, AU - Laffeber,Charlie, AU - Lebbink,Joyce H G, AU - Bruens,Serena, AU - Gaskin,Duncan, AU - Beerens,Dior, AU - Klunder,Maarten, AU - Joosten,Rob, AU - Demmers,Jeroen A A, AU - van Gent,Dik, AU - Mouton,Johan W, AU - van der Spek,Peter J, AU - van der Oost,John, AU - van Baarlen,Peter, AU - Louwen,Rogier, Y1 - 2020/06/17/ PY - 2019/09/12/received PY - 2020/05/06/accepted PY - 2020/6/30/entrez SP - eaaz4849 EP - eaaz4849 JF - Science advances JO - Sci Adv VL - 6 IS - 25 N2 - 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. SN - 2375-2548 UR - https://www.unboundmedicine.com/medline/citation/32596446/Guide-free_Cas9_from_pathogenic_Campylobacter_jejuni_bacteria_causes_severe_damage_to_DNA L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/32596446/ DB - PRIME DP - Unbound Medicine ER -
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