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.
References
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).
Salomon, W. E., Jolly, S. M., Moore, M. J., Zamore, P. D. & Serebrov, V. Cell 162, 84–95 (2015).
Ameres, S. L., Martinez, J. & Schroeder, R. Cell 130, 101–112 (2007).
Becker, W. R. et al. Mol. Cell 75, 741–755 e11 (2019).
McGeary, S. E. et al. Science 366, eaav1741 (2019).
Gong, S., Yu, H. H., Johnson, K. A. & Taylor, D. W. Cell Rep. 22, 359–371 (2018).
Singh, D., Sternberg, S. H., Fei, J., Doudna, J. A. & Ha, T. Nat. Commun. 7, 12778 (2016).
Raper, A. T., Stephenson, A. A. & Suo, Z. J. Am. Chem. Soc. 140, 2971–2984 (2018).
Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C. & Doudna, J. A. Nature 507, 62–67 (2014).
Shore, D., Langowski, J. & Baldwin, R. L. Proc. Natl Acad. Sci. USA 78, 4833–4837 (1981).
Kahn, J. D. & Crothers, D. M. Proc. Natl Acad. Sci. USA 89, 6343–6347 (1992).
Borowiec, J. A., Zhang, L., Sasse-Dwight, S. & Gralla, J. D. J. Mol. Biol. 196, 101–111 (1987).
Mekler, V., Minakhin, L. & Severinov, K. Proc. Natl Acad. Sci. USA 114, 5443–5448 (2017).
Strohkendl, I., Saifuddin, F. A., Rybarski, J. R., Finkelstein, I. J. & Russell, R. Mol. Cell 71, 816–824 e3 (2018).
Swarts, D. C., van der Oost, J. & Jinek, M. Mol. Cell 66, 221–233 e4 (2017).
Singh, D. et al. Proc. Natl Acad. Sci. USA 115, 5444–5449 (2018).
Bell, J. C. & Kowalczykowski, S. C. Trends Biochem. Sci. 41, 491–507 (2016).
Chen, Z., Yang, H. & Pavletich, N. P. Nature 453, 489–494 (2008).
Yang, H., Zhou, C., Dhar, A. & Pavletich, N. P. Nature 586, 801–806 (2020).
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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
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DOI: https://doi.org/10.1038/s41594-022-00763-1
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