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Single molecule analysis of combinatorial splicing.

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.

SUBMITTER: Conze T

PROVIDER: S-EPMC2938221 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature

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Single molecule analysis of combinatorial splicing.

Conze Tim T Göransson Jenny J Razzaghian Hamid Reza HR Ericsson Olle O Oberg Daniel D Akusjärvi Göran G Landegren Ulf U Nilsson Mats M

Nucleic acids research 20100629 16

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 ...[more]

PMID: 20587504

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