Figure 1.

A high-throughput screening strategy for regulators of alternative splicing. Two dual-fluorescence minigenes, pflareA (A) and pflareG (B), are constructed to report splicing of an alternative exon with opposite outputs. In the pre-mRNA schematics, exons in boxes are connected by horizontal lines representing introns. After splicing, the exon-included and exon-skipped mRNA isoforms contain different open reading frames (yellow highlight). Exon 18 is spliced at similar levels in the pflareA-exon 18 and the pflareG-exon 18 mRNAs (A,B). However, the pflareA-exon 18 reporter expresses low RFP and high GFP, whereas the pflareG-exon 18 reporter expresses high RFP and low GFP, upon transient expression in N2a cells. (C) The framework for the cell-based high-throughput screens to identify splicing regulatory factors.