A multiplexed homology-directed DNA repair assay reveals the impact of ~1,700 BRCA1 variants on protein function
| Autor: | Muhtadi M. Islam, Stanley Fields, Aleksandra I. Adamovich, Tapahsama Banerjee, Jeffrey D. Parvin, Jay Shendure, Lea M. Starita, Justin Gullingsrud |
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| Rok vydání: | 2018 |
| Předmět: |
0303 health sciences
Reporter gene endocrine system diseases medicine.diagnostic_test DNA repair Genomics Computational biology Biology Homology (biology) Homology directed repair 03 medical and health sciences 0302 clinical medicine 030220 oncology & carcinogenesis medicine Missense mutation Multiplex skin and connective tissue diseases 030304 developmental biology Genetic testing |
| DOI: | 10.1101/295279 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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