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A multiplexed homology-directed DNA repair assay reveals the impact of ~1,700 BRCA1 variants on protein function

Lea M. Starita, Muhtadi M. Islam, Tapahsama Banerjee, Aleksandra I. Adamovich, Justin Gullingsrud, Stanley Fields, Jay Shendure, Jeffrey D. Parvin
doi: https://doi.org/10.1101/295279
Lea M. Starita
1Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
2Brotman Baty Institute for Precision Medicine, Seattle, Washington, USA.
7These authors contributed equally to this work.
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Muhtadi M. Islam
3Department of Biomedical Informatics, The Ohio State University, Columbus, Ohio, USA.
7These authors contributed equally to this work.
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Tapahsama Banerjee
3Department of Biomedical Informatics, The Ohio State University, Columbus, Ohio, USA.
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Aleksandra I. Adamovich
3Department of Biomedical Informatics, The Ohio State University, Columbus, Ohio, USA.
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Justin Gullingsrud
1Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
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Stanley Fields
1Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
4Department of Medicine, University of Washington, Seattle, Washington, USA.
5Howard Hughes Medical Institute, Seattle, Washington, USA.
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Jay Shendure
1Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
2Brotman Baty Institute for Precision Medicine, Seattle, Washington, USA.
5Howard Hughes Medical Institute, Seattle, Washington, USA.
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  • For correspondence: Jeffrey.Parvin@osumc.edu shendure@u.washington.edu
Jeffrey D. Parvin
3Department of Biomedical Informatics, The Ohio State University, Columbus, Ohio, USA.
6The Ohio State University Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, USA.
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  • For correspondence: Jeffrey.Parvin@osumc.edu shendure@u.washington.edu
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Abstract

Loss-of-function mutations in BRCA1 confer a predisposition to breast and ovarian cancer. Genetic testing for mutations in the BRCA1 gene frequently reveals a missense variant for which the impact on the molecular function of the BRCA1 protein is unknown. Functional BRCA1 is required for homology directed repair (HDR) of double-strand DNA breaks, a key activity for maintaining genome integrity and tumor suppression. Here we describe a multiplex HDR reporter assay to simultaneously measure the effect of hundreds of variants of BRCA1 on its role in DNA repair. Using this assay, we measured the effects of ~1,700 amino acid substitutions in the first 302 residues of BRCA1. Benchmarking these results against variants with known effects, we demonstrate accurate discrimination of loss-of-function versus benign variants. We anticipate that this assay can be used to functionally characterize BRCA1 missense variants at scale, even before the variants are observed in results from genetic testing.

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Posted April 05, 2018.
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A multiplexed homology-directed DNA repair assay reveals the impact of ~1,700 BRCA1 variants on protein function
Lea M. Starita, Muhtadi M. Islam, Tapahsama Banerjee, Aleksandra I. Adamovich, Justin Gullingsrud, Stanley Fields, Jay Shendure, Jeffrey D. Parvin
bioRxiv 295279; doi: https://doi.org/10.1101/295279
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A multiplexed homology-directed DNA repair assay reveals the impact of ~1,700 BRCA1 variants on protein function
Lea M. Starita, Muhtadi M. Islam, Tapahsama Banerjee, Aleksandra I. Adamovich, Justin Gullingsrud, Stanley Fields, Jay Shendure, Jeffrey D. Parvin
bioRxiv 295279; doi: https://doi.org/10.1101/295279

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