Figure 1.
Outline of the proposed method. (A) Schematic representation of the reporter, which includes target gene fragment (an alternative exon with adjacent introns and constitutive exons) flanked by RFP- and GFP-coding regions. Alternative splicing results in two mRNA species. Translation of the normal full-length transcript (left) produces RFP and GFP, whereas translation of the alternative short transcript (right) results in RFP only. (B) Scheme of expected flow cytometry results (GFP-RFP bivariate plot). Gray area shows control cells expressing normal transcript without introns. Cells with only normal splicing, only alternative splicing and both types of splicing of the target gene are shown as yellow, red and orange dots, respectively. A percentage of normal and alternative transcripts in each cell can be estimated using designated equation.