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Rapid generation of splicing reporters with pSpliceExpress

2008; Elsevier BV; Volume: 427; Issue: 1-2 Linguagem: Inglês

10.1016/j.gene.2008.09.021

1879-0038

Shivendra Kishore, Amit Khanna, Stefan Stamm,

RNA and protein synthesis mechanisms

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.