Rapid generation of splicing reporters with pSpliceExpress

S Kishore, A Khanna, S Stamm - Gene, 2008 - Elsevier
Gene, 2008Elsevier
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 …
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.
Elsevier