Rapid generation of splicing reporters with pSpliceExpress

Shivendra Kishore, Amit Khanna, Stefan Stamm

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

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.

Original languageEnglish
Pages (from-to)104-110
Number of pages7
JournalGene
Volume427
Issue number1-2
DOIs
StatePublished - Dec 31 2008

Keywords

  • Alternative splicing
  • High throughput
  • Recombination
  • Reporter gene

ASJC Scopus subject areas

  • Genetics

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