Oct 22, 2008

Rapid generation of splicing reporters with pSpliceExpress

Gene
Shivendra KishoreStefan Stamm

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.

Mentioned in this Paper

Pre-mRNA Splicing
Polymerase
Transfection
Exons
Genome
Enzymes, antithrombotic
Primary RNA Transcript
Recombination, Genetic
DNA Sequence - Cloning Site
RAD51 wt Allele

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