Single molecule analysis of combinatorial splicing

Nucleic Acids Res. 2010 Sep;38(16):e163. doi: 10.1093/nar/gkq581. Epub 2010 Jun 29.

Abstract

Alternative splicing creates diverse mRNA isoforms from single genes and thereby enhances complexity of transcript structure and of gene function. We describe a method called spliceotyping, which translates combinatorial mRNA splicing patterns along transcripts into a library of binary strings of nucleic acid tags that encode the exon composition of individual mRNA molecules. The exon inclusion pattern of each analyzed transcript is thus represented as binary data, and the abundance of different splice variants is registered by counts of individual molecules. The technique is illustrated in a model experiment by analyzing the splicing patterns of the adenovirus early 1A gene and the beta actin reference transcript. The method permits many genes to be analyzed in parallel and it will be valuable for elucidating the complex effects of combinatorial splicing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / genetics
  • Adenovirus E1A Proteins / genetics
  • Alternative Splicing*
  • DNA Probes
  • HeLa Cells
  • Humans
  • Nucleic Acid Hybridization / methods*
  • RNA Splice Sites

Substances

  • Adenovirus E1A Proteins
  • DNA Probes
  • RNA Splice Sites