A bichromatic fluorescent reporter for cell-based screens of alternative splicing.

Abstract:

:Alternative splicing is the primary source of proteome complexity in metazoans and its regulation shapes the proteome in response to shifting physiological requirements. We developed a bichromatic splicing reporter that uses a peculiar feature of some fluorescent protein coding regions to express two different fluorescent proteins from a single alternative splicing event. The mutually exclusive expression of different fluorescent proteins from a single reporter provides a uniquely sensitive approach for high-throughput screening and analysis of cell-specific splicing events in mixed cell cultures and tissues of transgenic animals. This reporter is applicable to the majority of alternative splicing patterns and can be used to quantify alternative splicing within single cells and to select cells that express specific splicing patterns. The ability to perform quantitative single-cell analysis of alternative splicing and high-throughput screens will enhance progress toward understanding splicing regulatory networks and identifying compounds that reverse pathogenic splicing defects.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Orengo JP,Bundman D,Cooper TA

doi

10.1093/nar/gkl967

subject

Has Abstract

pub_date

2006-01-01 00:00:00

pages

e148

issue

22

eissn

0305-1048

issn

1362-4962

pii

gkl967

journal_volume

34

pub_type

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