Molecular characterization of ten F8 splicing mutations in RNA isolated from patient's leucocytes: assessment of in silico prediction tools accuracy

摘要: Although 8% of reported FVIII gene (F8) mutations responsible for haemophilia A (HA) affect mRNA processing, very few have been fully characterized at the level and/or systematically predicted their biological consequences by in silico analysis. This study is aimed to elucidate effect potential splice site (PSSM) on F8 investigate its correlation with disease severity, and assess concordance predictions. We studied from 10 HA patient's leucocytes PSSM RT-PCR compared experimental results those silico. The analysis could explain all phenotypes observed demonstrated exon skipping six cases (c.222G>A, c.601+1delG, c.602-11T>G, c.671-3C>G, c.6115+9C>G c.6116-1G>A) activation cryptic splicing sites, both donor (c.1009+1G>A c.1009+3A>C) acceptor sites (c.266-3delC c.5587-1G>A). In contrast, was able predict score variation most affected site, but precise mechanism only be correctly determined two analysed. addition, we detected aberrant transcripts, even healthy controls, so this must taken into account as they mask actual contribution some PSSM. conclude that using still constitutes an excellent approach transcriptional effects HA, whereas prediction not always reliable diagnostic decision-making.