In vitro coupled transcription splicing

Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here...

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Bibliographic Details
Published inMethods (San Diego, Calif.) Vol. 37; no. 4; pp. 314 - 322
Main Authors Natalizio, Barbara J., Garcia-Blanco, Mariano A.
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 01.12.2005
Subjects
Cotranscriptional splicing
Coupling
Deoxyribonucleases
DNA-Directed RNA Polymerases - genetics
Exons
Globins - biosynthesis
Humans
Molecular Biology - methods
mRNA processing
RNA Interference
RNA polymerase II
RNA Polymerase II - genetics
RNA Precursors - physiology
RNA Splicing
T7 RNA polymerase
Temperature
Transcription
Transcription, Genetic
Viral Proteins - genetics
T7 RNA polymerase
Cotranscriptional splicing
Coupling
Transcription
mRNA processing
RNA polymerase II
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Summary:Many convincing studies published in recent years strongly support coupling of transcription and pre-mRNA processing. Despite key advances in our understanding of these processes, there is a lack of a robust in vitro system in which to study the mechanism of this coupling for complex pre-mRNAs. Here, we describe an in vitro system capable of transcribing and splicing complex transcripts with three and four exons. We also demonstrate how the system can be used to study exon silencing in vitro. We believe that this system will be a useful tool to study the mechanisms that mediate the coupling of transcription and pre-mRNA processing.
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ISSN:1046-2023
1095-9130
DOI:10.1016/j.ymeth.2005.07.013