TY - JOUR
T1 - A Genetic Map of the Response to DNA Damage in Human Cells
AU - Olivieri, Michele
AU - Cho, Tiffany
AU - Álvarez-Quilón, Alejandro
AU - Li, Kejiao
AU - Schellenberg, Matthew J.
AU - Zimmermann, Michal
AU - Hustedt, Nicole
AU - Rossi, Silvia Emma
AU - Adam, Salomé
AU - Melo, Henrique
AU - Heijink, Anne Margriet
AU - Sastre-Moreno, Guillermo
AU - Moatti, Nathalie
AU - Szilard, Rachel K.
AU - McEwan, Andrea
AU - Ling, Alexanda K.
AU - Serrano-Benitez, Almudena
AU - Ubhi, Tajinder
AU - Feng, Sumin
AU - Pawling, Judy
AU - Delgado-Sainz, Irene
AU - Ferguson, Michael W.
AU - Dennis, James W.
AU - Brown, Grant W.
AU - Cortés-Ledesma, Felipe
AU - Williams, R. Scott
AU - Martin, Alberto
AU - Xu, Dongyi
AU - Durocher, Daniel
N1 - Funding Information:
We thank J.Y. Yuan for technical assistance as well as J. Moffat, S. Angers, and T. Hart for the TKO libraries and protocols and C. Boone for yeast strains. We also thank F. Meng, S. Rottenberg, and M. Luijsterburg for sharing unpublished results. A.A.Q. G.S.M. and A.M.H. are recipients of long-term EMBO fellowships (ALTF 910-2017, 795-2017, and 124-2019), S.E.R. was supported by a fellowship from AIRC, and S.A. was a Banting post-doctoral fellow. A.S.B. was supported by a PhD Studentship from Asociación Española Familia Ataxia Telangiectasia and an EMBO short-term fellowship. The ICRF-187 was funded by the Spanish Government (SAF2017-89619-R, European Regional Development Fund) and European Research Council (ERC-CoG-2014-647359). Work in the R.S.W. laboratory was supported in part by the US National Institutes of Health Intramural Program, US National Institute of Environmental Health Sciences (NIEHS, 1Z01ES102765). D.D. is a Canada Research Chair (Tier I), and the work was supported by grants from the CIHR (FDN143343 to D.D.; FRN 123518 and PJT-156330 to A.Martin) and Canadian Cancer Society grant (705644 to D.D.) with additional support to D.D. from the Krembil Foundation. Conceptualization, M.O. and D.D.; Methodology, M.O. A.A.-Q. M.J.S. J.P. J.W.D. R.S.W. and D.D.; Formal Analysis and Data Curation, H.M. and A. McEwan; Investigation, M.O. T.C. A.A.-Q. K.L. M.J.S. M.Z. N.H. S.E.R. S.A. A.M.H. G.S.-M. N.M. R.K.S. A.K.L. A.S.-B. T.U. S.F. J.P. I.D.-S. and M.W.F.; Writing – Original Draft, M.O. H.M. and D.D.; Writing – Reviewing and Editing, M.O. R.K.S. and D.D.; Visualization, M.O. A.A.-Q. H.M. and D.D.; Funding Acquisition, J.W.D. G.W.B. F.C.-L. R.S.W. A. Martin, D.X. and D.D.; Supervision, J.W.D. G.W.B. F.C.-L. R.S.W. A. Martin, D.X. and D.D. Michal Zimmermann is an employee and shareholder of Repare Therapeutics. Daniel Durocher is a founder of Repare Therapeutics and a member of its scientific advisory board.
Funding Information:
We thank J.Y. Yuan for technical assistance as well as J. Moffat, S. Angers, and T. Hart for the TKO libraries and protocols and C. Boone for yeast strains. We also thank F. Meng, S. Rottenberg, and M. Luijsterburg for sharing unpublished results. A.A.Q., G.S.M., and A.M.H. are recipients of long-term EMBO fellowships ( ALTF 910-2017 , 795-2017 , and 124-2019 ), S.E.R. was supported by a fellowship from AIRC , and S.A. was a Banting post-doctoral fellow. A.S.B. was supported by a PhD Studentship from Asociación Española Familia Ataxia Telangiectasia and an EMBO short-term fellowship. The ICRF-187 was funded by the Spanish Government ( SAF2017-89619-R , European Regional Development Fund) and European Research Council ( ERC-CoG-2014-647359 ). Work in the R.S.W. laboratory was supported in part by the US National Institutes of Health Intramural Program, US National Institute of Environmental Health Sciences (NIEHS, 1Z01ES102765 ). D.D. is a Canada Research Chair (Tier I), and the work was supported by grants from the CIHR ( FDN143343 to D.D.; FRN 123518 and PJT-156330 to A.Martin) and Canadian Cancer Society grant ( 705644 to D.D.) with additional support to D.D. from the Krembil Foundation .
Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020/7/23
Y1 - 2020/7/23
N2 - The response to DNA damage is critical for cellular homeostasis, tumor suppression, immunity, and gametogenesis. In order to provide an unbiased and global view of the DNA damage response in human cells, we undertook 31 CRISPR-Cas9 screens against 27 genotoxic agents in the retinal pigment epithelium-1 (RPE1) cell line. These screens identified 890 genes whose loss causes either sensitivity or resistance to DNA-damaging agents. Mining this dataset, we discovered that ERCC6L2 (which is mutated in a bone-marrow failure syndrome) codes for a canonical non-homologous end-joining pathway factor, that the RNA polymerase II component ELOF1 modulates the response to transcription-blocking agents, and that the cytotoxicity of the G-quadruplex ligand pyridostatin involves trapping topoisomerase II on DNA. This map of the DNA damage response provides a rich resource to study this fundamental cellular system and has implications for the development and use of genotoxic agents in cancer therapy.
AB - The response to DNA damage is critical for cellular homeostasis, tumor suppression, immunity, and gametogenesis. In order to provide an unbiased and global view of the DNA damage response in human cells, we undertook 31 CRISPR-Cas9 screens against 27 genotoxic agents in the retinal pigment epithelium-1 (RPE1) cell line. These screens identified 890 genes whose loss causes either sensitivity or resistance to DNA-damaging agents. Mining this dataset, we discovered that ERCC6L2 (which is mutated in a bone-marrow failure syndrome) codes for a canonical non-homologous end-joining pathway factor, that the RNA polymerase II component ELOF1 modulates the response to transcription-blocking agents, and that the cytotoxicity of the G-quadruplex ligand pyridostatin involves trapping topoisomerase II on DNA. This map of the DNA damage response provides a rich resource to study this fundamental cellular system and has implications for the development and use of genotoxic agents in cancer therapy.
KW - CRISPR
KW - DNA damage
KW - DNA repair
KW - DNA-damaging agents
KW - cancer therapeutics
KW - functional genomics
KW - genome stability
KW - mechanism-of-action
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U2 - 10.1016/j.cell.2020.05.040
DO - 10.1016/j.cell.2020.05.040
M3 - Article
C2 - 32649862
AN - SCOPUS:85087739045
SN - 0092-8674
VL - 182
SP - 481-496.e21
JO - Cell
JF - Cell
IS - 2
ER -