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Assessing kinetics and recruitment of DNA repair factors using high content screens.
Martinez-Pastor, Barbara; Silveira, Giorgia G; Clarke, Thomas L; Chung, Dudley; Gu, Yuchao; Cosentino, Claudia; Davidow, Lance S; Mata, Gadea; Hassanieh, Sylvana; Salsman, Jayme; Ciccia, Alberto; Bae, Narkhyun; Bedford, Mark T; Megias, Diego; Rubin, Lee L; Efeyan, Alejo; Dellaire, Graham; Mostoslavsky, Raul.
  • Martinez-Pastor B; The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA 02114, USA; Molecular Oncology Program, Spanish National Cancer Research Center (CNIO), Madrid 28029, Spain. Electronic address: bmartinezp@cnio.es.
  • Silveira GG; The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA 02114, USA; The Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
  • Clarke TL; The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA 02114, USA; The Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
  • Chung D; Department of Pathology, Dalhousie University, Halifax, NS B3H 4R2, Canada.
  • Gu Y; School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.
  • Cosentino C; The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA 02114, USA.
  • Davidow LS; Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA.
  • Mata G; Confocal Microscopy Unit, Spanish National Cancer Research Center (CNIO), Madrid 28029, Spain.
  • Hassanieh S; The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA 02114, USA.
  • Salsman J; Department of Pathology, Dalhousie University, Halifax, NS B3H 4R2, Canada.
  • Ciccia A; Department of Genetics and Development, Columbia University Medical Center, New York, NY 10032, USA; Herbert Irving Comprehensive Cancer Center, Columbia University Medical Center, New York, NY 10032, USA.
  • Bae N; Department of Genetics and Development, Columbia University Medical Center, New York, NY 10032, USA.
  • Bedford MT; Department of Epigenetics & Molecular Carcinogenesis, M.D.Anderson Cancer Center, University of Texas, Smithville, TX 78957, USA.
  • Megias D; Confocal Microscopy Unit, Spanish National Cancer Research Center (CNIO), Madrid 28029, Spain.
  • Rubin LL; Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA.
  • Efeyan A; Molecular Oncology Program, Spanish National Cancer Research Center (CNIO), Madrid 28029, Spain.
  • Dellaire G; Department of Pathology, Dalhousie University, Halifax, NS B3H 4R2, Canada; Beatrice Hunter Cancer Research Institute, Halifax, NS B3H 4R2, Canada. Electronic address: dellaire@dal.ca.
  • Mostoslavsky R; The Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA 02114, USA; The Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA. Electronic address: rmostoslavsky@mgh.harvard.edu.
Cell Rep ; 37(13): 110176, 2021 12 28.
Article en En | MEDLINE | ID: mdl-34965416
ABSTRACT
Repair of genetic damage is coordinated in the context of chromatin, so cells dynamically modulate accessibility at DNA breaks for the recruitment of DNA damage response (DDR) factors. The identification of chromatin factors with roles in DDR has mostly relied on loss-of-function screens while lacking robust high-throughput systems to study DNA repair. In this study, we have developed two high-throughput systems that allow the study of DNA repair kinetics and the recruitment of factors to double-strand breaks in a 384-well plate format. Using a customized gain-of-function open-reading frame library ("ChromORFeome" library), we identify chromatin factors with putative roles in the DDR. Among these, we find the PHF20 factor is excluded from DNA breaks, affecting DNA repair by competing with 53BP1 recruitment. Adaptable for genetic perturbations, small-molecule screens, and large-scale analysis of DNA repair, these resources can aid our understanding and manipulation of DNA repair.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Daño del ADN / Cromatina / Histonas / Sistemas de Lectura Abierta / Enzimas Reparadoras del ADN / Reparación del ADN / Proteína 1 de Unión al Supresor Tumoral P53 Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Año: 2021 Tipo del documento: Article
Texto completo
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PubMed Links
Search on Google

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Daño del ADN / Cromatina / Histonas / Sistemas de Lectura Abierta / Enzimas Reparadoras del ADN / Reparación del ADN / Proteína 1 de Unión al Supresor Tumoral P53 Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Año: 2021 Tipo del documento: Article