Rapid generation of splicing reporters with pSpliceExpress

Gene. 2008 Dec 31;427(1-2):104-10. doi: 10.1016/j.gene.2008.09.021. Epub 2008 Oct 1.

Abstract

Almost all human protein-coding transcripts undergo pre-mRNA splicing and a majority of them is alternatively spliced. The most common technique used to analyze the regulation of an alternative exon is through reporter minigene constructs. However, their construction is time-consuming and is often complicated by the limited availability of appropriate restriction sites. Here, we report a fast and simple recombination-based method to generate splicing reporter genes, using a new vector, pSpliceExpress. The system allows generation of minigenes within one week. Minigenes generated with pSpliceExpress show the same regulation as displayed by conventionally cloned reporter constructs and provide an alternate avenue to study splice site selection in vivo.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • DNA Restriction Enzymes / chemistry
  • Exons
  • Genes, Reporter
  • Genetic Techniques*
  • Humans
  • Models, Genetic
  • Molecular Sequence Data
  • RNA Precursors / genetics
  • RNA Splicing*
  • RNA, Messenger / metabolism
  • Recombination, Genetic

Substances

  • RNA Precursors
  • RNA, Messenger
  • DNA Restriction Enzymes