Guide-free Cas9 from pathogenic <i>Campylobacter jejuni</i> bacteria causes severe damage to DNA

Chinmoy Saha, Prarthana Mohanraju, Andrew Stubbs, Gaurav Dugar, Youri Hoogstrate, Gert‐Jan Kremers, Wiggert A. van Cappellen, Deborah Horst-Kreft, Charlie Laffeber, Joyce H.G. Lebbink,

Science Advances · 2020 · 45 citations · 38 references

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Abstract

(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.

References

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