Figure 4
Detection of COL7A1-RTM fusion transcripts by sqRT-PCR. (a) Schematic depiction of the endogenous RTM28, containing a FLAG tag and silent mutations within the wild-type coding domain of COL7A1. (b) SqRT-PCR analysis, performed on total RNA isolated from RTM28-transduced RDEB keratinocytes using primers spanning exons 46–49, revealed the expression of COL7A1-RTM fusion transcripts at the RNA level. The respective PCR product with a size of 205 bp was verified by sequence analysis. (c) SqRT-PCR was performed to quantify total COL7A1 transcripts present in RTM-treated keratinocytes. As a result, a fivefold increase of COL7A1 expression was obtained after RTM28 treatment in comparison with RDEB patient cells transduced with an empty retroviral vector (RDEB-mock).
