Validation of a cell-based luciferase system for measurement of exon 10 splicing in the tau gene (adapted from Yu et al. ). (A) The luciferase mini-gene construct. The firefly luciferase gene was fused downstream of exon 11 in frame with a SI9/LI10 mini-gene containing the M14 (G to T) mutation. A stop codon was introduced into exon 10 by mutagenizing position 73 in exon 10 (A to T). The nucleotide A of the initiation codon ATG in the luciferase gene was converted to T. (B) A control mini-gene with a fused Renilla luciferase to exon 11. (C) The firefly luciferase mini-gene (A) was co-transfected into SKN-MC cells with constructs of SRp40, SRp20, or SRp55. Relative firefly luciferase activities were normalized using the Renilla luciferase control (B). (D) RT-PCR from cells described in (C) was carried out to quantify exon 10 inclusion/exclusion.
Zhou et al. BMC Neuroscience 2008 9(Suppl 2):S10 doi:10.1186/1471-2202-9-S2-S10