Assaying the Effects of Splice Site Variants by Exon Trapping in a Mammalian Cell Line.

PubMed ID: 28758139

Author(s): Tompson SW, Young TL. Assaying the effects of splice site variants by exon trapping in a mammalian cell line. Bio Protoc. 2017 May 20;7(10). pii: e2281. doi: 10.21769/BioProtoc.2281. PMID 28758139

Journal: Bio Protocol, Volume 7, Issue 10, May 2017

There are several in silico programs that endeavor to predict the functional impact of an individual’s sequence variation at splice donor/acceptor sites, but experimental confirmation is problematic without a source of RNA from the individual that carries the variant. With the aid of an exon trapping vector, such as pSPL3, an investigator can test whether a splice site sequence change leads to altered RNA splicing, through expression of reference and variant mini-genes in mammalian cells and analysis of the resultant RNA products.