Using mini-genes to identify factors that modulate alternative splicing

Methods Mol Biol. 2012:867:349-62. doi: 10.1007/978-1-61779-767-5_22.

Abstract

Many genetic mutations result in the disruption of (alternative) splicing. Prime examples are the SMN1 and SMN2 genes: a silent mutation in SMN2 leads to the skipping of the constitutive exon 7 in the majority of SMN2 transcripts, while this exon is generally included in SMN1 transcripts. Lack of SMN is embryonic lethal and loss of SMN1 genes leads to a severe decrease in SMN protein and is associated with spinal muscular atrophy. There are proteins and drugs that can chance alternative splicing events, e.g. increase the inclusion of exon 7 in SMN2. This chapter describes mini-genes and methods that can be employed to screen for candidate proteins and drugs.

MeSH terms

  • Alternative Splicing* / drug effects
  • Animals
  • Cell Line
  • Cloning, Molecular / methods
  • DNA / genetics
  • DNA / isolation & purification
  • DNA, Complementary / genetics
  • Humans
  • Muscular Atrophy, Spinal / drug therapy
  • Muscular Atrophy, Spinal / genetics*
  • Polymerase Chain Reaction / methods
  • RNA / genetics
  • RNA / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Survival of Motor Neuron 1 Protein / genetics*
  • Survival of Motor Neuron 2 Protein / genetics*
  • Transfection

Substances

  • DNA, Complementary
  • Survival of Motor Neuron 1 Protein
  • Survival of Motor Neuron 2 Protein
  • RNA
  • DNA