A High-Throughput Functional Complementation Assay for Classification ofBRCA1Missense Variants
Open Access
- 1 October 2013
- journal article
- Published by American Association for Cancer Research (AACR) in Cancer Discovery
- Vol. 3 (10), 1142-1155
- https://doi.org/10.1158/2159-8290.cd-13-0094
Abstract
Mutations in BRCA1 and BRCA2 account for the majority of hereditary breast and ovarian cancers, and therefore sequence analysis of both genes is routinely conducted in patients with early-onset breast cancer. Besides mutations that clearly abolish protein function or are known to increase cancer risk, a large number of sequence variants of uncertain significance (VUS) have been identified. Although several functional assays for BRCA1 VUSs have been described, thus far it has not been possible to conduct a high-throughput analysis in the context of the full-length protein. We have developed a relatively fast and easy cDNA-based functional assay to classify BRCA1 VUSs based on their ability to functionally complement BRCA1-deficient mouse embryonic stem cells. Using this assay, we have analyzed 74 unclassified BRCA1 missense mutants for which all predicted pathogenic variants are confined to the BRCA1 RING and BRCT domains.Keywords
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This publication has 48 references indexed in Scilit:
- A guide for functional analysis ofBRCA1variants of uncertain significanceHuman Mutation, 2012
- Guidelines for splicing analysis in molecular diagnosis derived from a set of 327 combined in silico/in vitro studies on BRCA1 and BRCA2 variantsHuman Mutation, 2012
- BRCA1 Functions Independently of Homologous Recombination in DNA Interstrand Crosslink RepairMolecular Cell, 2012
- ENIGMA-Evidence-based network for the interpretation of germline mutant alleles: An international initiative to evaluate risk and clinical significance associated with sequence variation in BRCA1 and BRCA2 genesHuman Mutation, 2011
- A review of a multifactorial probability-based model for classification of BRCA1 and BRCA2 variants of uncertain significance (VUS)Human Mutation, 2011
- BRCA1 tumour suppression occurs via heterochromatin-mediated silencingNature, 2011
- 53BP1 Inhibits Homologous Recombination in Brca1-Deficient Cells by Blocking Resection of DNA BreaksCell, 2010
- A Selective Requirement for 53BP1 in the Biological Response to Genomic Instability Induced by Brca1 DeficiencyMolecular Cell, 2009
- Mouse embryonic stem cell–based functional assay to evaluate mutations in BRCA2Nature Medicine, 2008
- Evaluation of in silico splice tools for decision-making in molecular diagnosisHuman Mutation, 2008