Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening

Research output: Contribution to journal › Journal article › Research › peer-review

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Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening. / Lee, Sang-Woo; Tran, Kim Tai; Vazquez-Uribe, Ruben; Gotfredsen, Charlotte Held; Clausen, Mads Hartvig; Mendez, Blanca Lopez; Montoya, Guillermo; Bach, Anders; Sommer, Morten Otto Alexander.

In: Journal of Medicinal Chemistry, Vol. 65, No. 4, 2022, p. 3266–3305.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Lee, S-W, Tran, KT, Vazquez-Uribe, R, Gotfredsen, CH, Clausen, MH, Mendez, BL, Montoya, G, Bach, A & Sommer, MOA 2022, 'Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening', Journal of Medicinal Chemistry, vol. 65, no. 4, pp. 3266–3305. https://doi.org/10.1021/acs.jmedchem.1c01834

APA

Lee, S-W., Tran, K. T., Vazquez-Uribe, R., Gotfredsen, C. H., Clausen, M. H., Mendez, B. L., Montoya, G., Bach, A., & Sommer, M. O. A. (2022). Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening. Journal of Medicinal Chemistry, 65(4), 3266–3305. https://doi.org/10.1021/acs.jmedchem.1c01834

Vancouver

Lee S-W, Tran KT, Vazquez-Uribe R, Gotfredsen CH, Clausen MH, Mendez BL et al. Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening. Journal of Medicinal Chemistry. 2022;65(4):3266–3305. https://doi.org/10.1021/acs.jmedchem.1c01834

Author

Lee, Sang-Woo ; Tran, Kim Tai ; Vazquez-Uribe, Ruben ; Gotfredsen, Charlotte Held ; Clausen, Mads Hartvig ; Mendez, Blanca Lopez ; Montoya, Guillermo ; Bach, Anders ; Sommer, Morten Otto Alexander. / Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening. In: Journal of Medicinal Chemistry. 2022 ; Vol. 65, No. 4. pp. 3266–3305.

Bibtex

@article{2bc1cc566ac044bea6482689fd06a923,
title = "Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening",
abstract = "CRISPR/Cas9 has revolutionized several areas of life science; however, methods to control the Cas9 activity are needed for both scientific and therapeutic applications. Anti-CRISPR proteins are known to inhibit the CRISPR/Cas adaptive immunity; however, in vivo delivery of such proteins is problematic. Instead, small-molecule Cas9 inhibitors could serve as useful tools due to their permeable, proteolytically stable, and non-immunogenic nature. Here, we identified a small-molecule ligand with anti-CRISPR/Cas9 activity through a high-throughput screening utilizing an Escherichia coli selection system. Extensive structure–activity relationship studies, which involved a deconstruction–reconstruction strategy, resulted in a range of analogues with significant improvements in the inhibitory activity. Based on NMR and electrophoretic mobility shift assays, we propose that the inhibitory action of these compounds likely results from direct binding to apo-Cas9, preventing Cas9:gRNA complex formation. These molecules may find use as Cas9 modulators in various applications.",
author = "Sang-Woo Lee and Tran, {Kim Tai} and Ruben Vazquez-Uribe and Gotfredsen, {Charlotte Held} and Clausen, {Mads Hartvig} and Mendez, {Blanca Lopez} and Guillermo Montoya and Anders Bach and Sommer, {Morten Otto Alexander}",
year = "2022",
doi = "10.1021/acs.jmedchem.1c01834",
language = "English",
volume = "65",
pages = "3266–3305",
journal = "Journal of Medicinal Chemistry",
issn = "0022-2623",
publisher = "American Chemical Society",
number = "4",

}

RIS

TY - JOUR

T1 - Identification and Optimization of Novel Small-Molecule Cas9 Inhibitors by Cell-Based High-Throughput Screening

AU - Lee, Sang-Woo

AU - Tran, Kim Tai

AU - Vazquez-Uribe, Ruben

AU - Gotfredsen, Charlotte Held

AU - Clausen, Mads Hartvig

AU - Mendez, Blanca Lopez

AU - Montoya, Guillermo

AU - Bach, Anders

AU - Sommer, Morten Otto Alexander

PY - 2022

Y1 - 2022

N2 - CRISPR/Cas9 has revolutionized several areas of life science; however, methods to control the Cas9 activity are needed for both scientific and therapeutic applications. Anti-CRISPR proteins are known to inhibit the CRISPR/Cas adaptive immunity; however, in vivo delivery of such proteins is problematic. Instead, small-molecule Cas9 inhibitors could serve as useful tools due to their permeable, proteolytically stable, and non-immunogenic nature. Here, we identified a small-molecule ligand with anti-CRISPR/Cas9 activity through a high-throughput screening utilizing an Escherichia coli selection system. Extensive structure–activity relationship studies, which involved a deconstruction–reconstruction strategy, resulted in a range of analogues with significant improvements in the inhibitory activity. Based on NMR and electrophoretic mobility shift assays, we propose that the inhibitory action of these compounds likely results from direct binding to apo-Cas9, preventing Cas9:gRNA complex formation. These molecules may find use as Cas9 modulators in various applications.

AB - CRISPR/Cas9 has revolutionized several areas of life science; however, methods to control the Cas9 activity are needed for both scientific and therapeutic applications. Anti-CRISPR proteins are known to inhibit the CRISPR/Cas adaptive immunity; however, in vivo delivery of such proteins is problematic. Instead, small-molecule Cas9 inhibitors could serve as useful tools due to their permeable, proteolytically stable, and non-immunogenic nature. Here, we identified a small-molecule ligand with anti-CRISPR/Cas9 activity through a high-throughput screening utilizing an Escherichia coli selection system. Extensive structure–activity relationship studies, which involved a deconstruction–reconstruction strategy, resulted in a range of analogues with significant improvements in the inhibitory activity. Based on NMR and electrophoretic mobility shift assays, we propose that the inhibitory action of these compounds likely results from direct binding to apo-Cas9, preventing Cas9:gRNA complex formation. These molecules may find use as Cas9 modulators in various applications.

U2 - 10.1021/acs.jmedchem.1c01834

DO - 10.1021/acs.jmedchem.1c01834

M3 - Journal article

C2 - 35142491

VL - 65

SP - 3266

EP - 3305

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

SN - 0022-2623

IS - 4

ER -

ID: 291871793