Exploiting HSD17B11-dependent dialkynylcarbinols cytotoxicity for facile CRISPR/Cas9-based gene inactivation
Abstract
Several approaches have been developed to improve the efficiency of CRISPR/Cas9-based genome editing, including the co-inactivation of a gene whose loss confers resistance to a cytotoxic compound, thereby enabling enrichment of successfully edited cells. Here, we show across multiple cell lines that inactivation of HSD17B11, a non-essential member of the Short-chain Dehydrogenase/Reductase (SDR) superfamily, confers strong resistance (29-to 131-fold) to a Phenyl diAlkynylCarbinol compound (PAC) in both human and mouse cells, without affecting cell viability or proliferation. We demonstrate that co-inactivation of HSD17B11 followed by PAC selection can be used to rapidly identify efficient guide RNAs targeting a gene of interest and to readily isolate clones inactivated for one or multiple genes. Altogether, these results establish a simple and efficient experimental strategy for generating knockout cells by using PAC selection to enrich for successfully edited cells.
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BibTeX
@misc{dumais2026exploiting,
title = {{Exploiting HSD17B11-dependent dialkynylcarbinols cytotoxicity for facile CRISPR/Cas9-based gene inactivation}},
author = {Bastien Dumais and Madeleine Bossaert and Patrick Seigneur and Alexandrine Rozié and Sophie Gasmi and Luce Fontanié--Roger and Maëlle Caroff and Valérie Maraval and Vania Bernardes-Génisson and Dennis Gomez and Philippe Frit and Stéphanie Ballereau and Yves Génisson and Sébastien Britton},
howpublished = {HAL},
year = {2026},
month = may,
doi = {10.64898/2026.05.13.724824},
url = {https://cnrs.hal.science/hal-05772769v1},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - UNPB
TI - Exploiting HSD17B11-dependent dialkynylcarbinols cytotoxicity for facile CRISPR/Cas9-based gene inactivation
AU - Bastien Dumais
AU - Madeleine Bossaert
AU - Patrick Seigneur
AU - Alexandrine Rozié
AU - Sophie Gasmi
AU - Luce Fontanié--Roger
AU - Maëlle Caroff
AU - Valérie Maraval
AU - Vania Bernardes-Génisson
AU - Dennis Gomez
AU - Philippe Frit
AU - Stéphanie Ballereau
AU - Yves Génisson
AU - Sébastien Britton
PY - 2026
DA - 2026/05/13
PB - HAL
DO - 10.64898/2026.05.13.724824
UR - https://cnrs.hal.science/hal-05772769v1
AB - Several approaches have been developed to improve the efficiency of CRISPR/Cas9-based genome editing, including the co-inactivation of a gene whose loss confers resistance to a cytotoxic compound, thereby enabling enrichment of successfully edited cells. Here, we show across multiple cell lines that inactivation of HSD17B11, a non-essential member of the Short-chain Dehydrogenase/Reductase (SDR) superfamily, confers strong resistance (29-to 131-fold) to a Phenyl diAlkynylCarbinol compound (PAC) in both human and mouse cells, without affecting cell viability or proliferation. We demonstrate that co-inactivation of HSD17B11 followed by PAC selection can be used to rapidly identify efficient guide RNAs targeting a gene of interest and to readily isolate clones inactivated for one or multiple genes. Altogether, these results establish a simple and efficient experimental strategy for generating knockout cells by using PAC selection to enrich for successfully edited cells.
ER -