I am studying the functional role of a specific gene that was found to be colocalized with the splicing factor SC-35. The next step is to investigate wether this gene really modulates alternative splicing. Is there any good protocol to perform an in vitro splicing assay?
Is the gene or the gene product co-localized with SC-35? Anyway, there are plenty of published protocols, but optimal conditions may vary considerably from one pre-mRNA to another. What is your target RNA?
Sure...splicing should be analysed by RT-PCR-based issues and bindung of SR proteins could be analyzed via adequate plasmid constructs and reporter assays.
e.g. see
Chandradas S et al., J Leukoc Biol. 2010 Jan;87(1):147-52. doi: 10.1189/jlb.0409252.
and/or
Tardos JG et al., J Thromb Haemost. 2008 May;6(5):877-84. doi: 10.1111/j.1538-7836.2008.02946.x.
and/or
Eisenreich A et al., Circ Res. 2009 Mar 13;104(5):589-99. doi: 10.1161/CIRCRESAHA.108.183905.
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