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Inhibition of CRISPR-Cas9 ribonucleoprotein complex assembly by anti-CRISPR AcrIIC2.

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Thavalingam A, Cheng Z, Garcia B, Huang X, Shah M, Sun W, Wang M, Harrington L, Hwang S, Hidalgo-Reyes Y, Sontheimer EJ, Doudna J, Davidson AR, Moraes TF, Wang Y, Maxwell KL
OpenAccess logo Nat Commun 10 2806 (2019)
Related entries: 6jd7, 6jdj, 6n05

Cited: 29 times
EuropePMC logo PMID: 31243272

Abstract

CRISPR-Cas adaptive immune systems function to protect bacteria from invasion by foreign genetic elements. The CRISPR-Cas9 system has been widely adopted as a powerful genome-editing tool, and phage-encoded inhibitors, known as anti-CRISPRs, offer a means of regulating its activity. Here, we report the crystal structures of anti-CRISPR protein AcrIIC2Nme alone and in complex with Nme1Cas9. We demonstrate that AcrIIC2Nme inhibits Cas9 through interactions with the positively charged bridge helix, thereby preventing sgRNA loading. In vivo phage plaque assays and in vitro DNA cleavage assays show that AcrIIC2Nme mediates its activity through a large electronegative surface. This work shows that anti-CRISPR activity can be mediated through the inhibition of Cas9 complex assembly.

Articles - 6jdx mentioned but not cited (1)

  1. Inhibition of CRISPR-Cas9 ribonucleoprotein complex assembly by anti-CRISPR AcrIIC2. Thavalingam A, Cheng Z, Garcia B, Huang X, Shah M, Sun W, Wang M, Harrington L, Hwang S, Hidalgo-Reyes Y, Sontheimer EJ, Doudna J, Davidson AR, Moraes TF, Wang Y, Maxwell KL. Nat Commun 10 2806 (2019)


Reviews citing this publication (12)

  1. Anti-CRISPR protein applications: natural brakes for CRISPR-Cas technologies. Marino ND, Pinilla-Redondo R, Csörgő B, Bondy-Denomy J. Nat Methods 17 471-479 (2020)
  2. Anti-CRISPRs: Protein Inhibitors of CRISPR-Cas Systems. Davidson AR, Lu WT, Stanley SY, Wang J, Mejdani M, Trost CN, Hicks BT, Lee J, Sontheimer EJ. Annu Rev Biochem 89 309-332 (2020)
  3. Controlling and enhancing CRISPR systems. Shivram H, Cress BF, Knott GJ, Doudna JA. Nat Chem Biol 17 10-19 (2021)
  4. Structures and Strategies of Anti-CRISPR-Mediated Immune Suppression. Wiegand T, Karambelkar S, Bondy-Denomy J, Wiedenheft B. Annu Rev Microbiol 74 21-37 (2020)
  5. CRISPR-Cas Systems and the Paradox of Self-Targeting Spacers. Wimmer F, Beisel CL. Front Microbiol 10 3078 (2019)
  6. Structure-based functional mechanisms and biotechnology applications of anti-CRISPR proteins. Jia N, Patel DJ. Nat Rev Mol Cell Biol 22 563-579 (2021)
  7. Alternative functions of CRISPR-Cas systems in the evolutionary arms race. Mohanraju P, Saha C, van Baarlen P, Louwen R, Staals RHJ, van der Oost J. Nat Rev Microbiol 20 351-364 (2022)
  8. Type II anti-CRISPR proteins as a new tool for synthetic biology. Zhang Y, Marchisio MA. RNA Biol 18 1085-1098 (2021)
  9. Unveil the Secret of the Bacteria and Phage Arms Race. Wang Y, Fan H, Tong Y. Int J Mol Sci 24 4363 (2023)
  10. Mechanisms regulating the CRISPR-Cas systems. Zakrzewska M, Burmistrz M. Front Microbiol 14 1060337 (2023)
  11. Recent advances in the use of CRISPR/Cas for understanding the early development of molecular gaps in glial cells. Barragán-Álvarez CP, Flores-Fernandez JM, Hernández-Pérez OR, Ávila-Gónzalez D, Díaz NF, Padilla-Camberos E, Dublan-García O, Gómez-Oliván LM, Diaz-Martinez NE. Front Cell Dev Biol 10 947769 (2022)
  12. The Many (Inter)faces of Anti-CRISPRs: Modulation of CRISPR-Cas Structure and Dynamics by Mechanistically Diverse Inhibitors. Belato HB, Lisi GP. Biomolecules 13 264 (2023)

Articles citing this publication (16)



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