Figure 1
Validation of the germline TP53 variants c.275 + 6T > C and c.376-2dupA by Sanger sequencing (a) and RNA level characterizations of TP53 splice variants (b) Agarose gel electrophoresis of the RT-PCR products of the TP53:c.375 + 6T > C variant carrier and controls. A very faint, 200 bp-shorter extra band is detectable at the c.375 + 6T > C variant carrier sample (c) Sanger sequencing of the RT-PCR product of the TP53:c.375 + 6T > C variant carrier. The extremely low level of aberrant splice product of the variant carrier, generated by weakening the canonical splice donor site and applying a pre-existing cryptic exonic donor GT site (shown with red bracket, chromosome position is indicated) (d) Screenshot of the TP53 exon 4 genomic region (hg 19) visualized by UCSC genome browser (https://genome-euro.ucsc.edu). The orange arrow indicates the position of the activated donor GT splice site, purple arrow marks the variant position. The measure of the aberrant splicing, although detectable, is extremely low, therefore negligible from a clinical point of view.
