Nucleic Acids Research Advance Access originally published online on November 16, 2006
Nucleic Acids Research 2006 34(22):e148; doi:10.1093/nar/gkl967
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Nucleic Acids Research, 2006, Vol. 34, No. 22 e148
© 2006 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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A bichromatic fluorescent reporter for cell-based screens of alternative splicing
1 Department of Pathology, Baylor College of Medicine Houston, TX 77030, USA 2 Department of Molecular and Cellular Biology, Baylor College of Medicine Houston, TX 77030, USA
*To whom correspondence should be addressed. Tel: +1 713 798 3141; Fax: +1 713 798 5838; Email: tcooper{at}bcm.tmc.edu
Received September 26, 2006. Revised October 16, 2006. Accepted October 17, 2006.
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.
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