ArticleNucleic acids research1988
Use of RNase H and primer extension to analyze RNA splicing.
Article in Nucleic acids research, 1988. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Alternative splicing of a human alpha-tropomyosin muscle-specific exon: identification of determining sequences.Molecular and cellular biology · 1992Article
- RNA processing is a limiting step for murine tumor necrosis factor beta expression in response to interleukin-2.Molecular and cellular biology · 1990Article
- Exon as well as intron sequences are cis-regulating elements for the mutually exclusive alternative splicing of the beta tropomyosin gene.Molecular and cellular biology · 1990Article
- A subfragment of the beta tropomyosin gene is alternatively spliced when transfected into differentiating muscle cells.Nucleic acids research · 1989Article
- Branch point selection in alternative splicing of tropomyosin pre-mRNAs.Nucleic acids research · 1989Article
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4 authors.
Funding
Abstract
A new method for the characterization of pre-mRNA splicing products is presented. In this method RNA molecules are hybridized to an oligodeoxynucleotide complementary to exon sequences upstream of a given 5' splice site, and the RNA strands of the resulting RNA:DNA hybrids are cleaved by RNase H. The cleaved RNAs are then subjected to primer extension using a 32P-labelled primer complementary to exon sequences downstream of an appropriate 3' splice site. Since the primer extension products all terminate at the site of RNase H cleavage, their lengths are indicative of the splice sites utilized. The method simplifies the study of the processing of complex pre-mRNAs by allowing the splicing events between any two exons to be analyzed. We have used this approach to characterize the RNAs generated by expression of the rat tropomyosin 1 (Tm 1) gene in various rat tissues and in cultured cells after transient transfection. The results demonstrate that this method is suitable for the analysis of alternative RNA processing in vivo.
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