Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • News & Views
  • Published:

RNA-GUIDED ENDONUCLEASES

A tweak and a peek: How Cas9 pries open double-stranded DNA to check its sequence

The CRISPR–Cas enzyme Cas9 faces the challenge of identifying a specific nucleotide sequence within double-stranded DNA. New cryo-EM and biochemical studies show that in the earliest steps of binding, Cas9 bends the DNA and promotes unwinding of two base pairs, enabling it to efficiently scan the sequence of this critical region.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Bending and unwinding of target DNA by Cas9.

References

  1. Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales, E. & Doudna, J. A. Nat. Struct. Mol. Biol. https://doi.org/10.1038/s41594-022-00756-0 (2022).

    Article  Google Scholar 

  2. Salomon, W. E., Jolly, S. M., Moore, M. J., Zamore, P. D. & Serebrov, V. Cell 162, 84–95 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Ameres, S. L., Martinez, J. & Schroeder, R. Cell 130, 101–112 (2007).

    Article  CAS  PubMed  Google Scholar 

  4. Becker, W. R. et al. Mol. Cell 75, 741–755 e11 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. McGeary, S. E. et al. Science 366, eaav1741 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Gong, S., Yu, H. H., Johnson, K. A. & Taylor, D. W. Cell Rep. 22, 359–371 (2018).

    Article  CAS  PubMed  Google Scholar 

  7. Singh, D., Sternberg, S. H., Fei, J., Doudna, J. A. & Ha, T. Nat. Commun. 7, 12778 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Raper, A. T., Stephenson, A. A. & Suo, Z. J. Am. Chem. Soc. 140, 2971–2984 (2018).

    Article  CAS  PubMed  Google Scholar 

  9. Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C. & Doudna, J. A. Nature 507, 62–67 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Shore, D., Langowski, J. & Baldwin, R. L. Proc. Natl Acad. Sci. USA 78, 4833–4837 (1981).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Kahn, J. D. & Crothers, D. M. Proc. Natl Acad. Sci. USA 89, 6343–6347 (1992).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Borowiec, J. A., Zhang, L., Sasse-Dwight, S. & Gralla, J. D. J. Mol. Biol. 196, 101–111 (1987).

    Article  CAS  PubMed  Google Scholar 

  13. Mekler, V., Minakhin, L. & Severinov, K. Proc. Natl Acad. Sci. USA 114, 5443–5448 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Strohkendl, I., Saifuddin, F. A., Rybarski, J. R., Finkelstein, I. J. & Russell, R. Mol. Cell 71, 816–824 e3 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Swarts, D. C., van der Oost, J. & Jinek, M. Mol. Cell 66, 221–233 e4 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Singh, D. et al. Proc. Natl Acad. Sci. USA 115, 5444–5449 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Bell, J. C. & Kowalczykowski, S. C. Trends Biochem. Sci. 41, 491–507 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Chen, Z., Yang, H. & Pavletich, N. P. Nature 453, 489–494 (2008).

    Article  CAS  PubMed  Google Scholar 

  19. Yang, H., Zhou, C., Dhar, A. & Pavletich, N. P. Nature 586, 801–806 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rick Russell.

Ethics declarations

Competing interests

The authors declare no competing interests.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sinan, S., Russell, R. A tweak and a peek: How Cas9 pries open double-stranded DNA to check its sequence. Nat Struct Mol Biol 29, 286–288 (2022). https://doi.org/10.1038/s41594-022-00763-1

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41594-022-00763-1

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing