Comprehensive splicing functional analysis of DNA variants of the BRCA2 gene by hybrid minigenes.

ABSTRACT:

Introduction

The underlying pathogenic mechanism of a large fraction of DNA variants of disease-causing genes is the disruption of the splicing process. We aimed to investigate the effect on splicing of the BRCA2 variants c.8488-1G > A (exon 20) and c.9026_9030del (exon 23), as well as 41 BRCA2 variants reported in the Breast Cancer Information Core (BIC) mutation database.

Methods

DNA variants were analyzed with the splicing prediction programs NNSPLICE and Human Splicing Finder. Functional analyses of candidate variants were performed by lymphocyte RT-PCR and/or hybrid minigene assays. Forty-one BIC variants of exons 19, 20, 23 and 24 were bioinformatically selected and generated by PCR-mutagenesis of the wild type minigenes.

Results

Lymphocyte RT-PCR of c.8488-1G >

SUBMITTER: Acedo A 

PROVIDER: S-EPMC3446350 | biostudies-literature | 2012 May

REPOSITORIES: biostudies-literature