Bio-Protocol (May 2017)

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

  • Stuart Tompson,
  • Terri Young

DOI
https://doi.org/10.21769/BioProtoc.2281
Journal volume & issue
Vol. 7, no. 10

Abstract

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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.