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Two-Color Fluorescent Reporters for Analysis of Alternative Splicing

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Alternative Splicing

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2537))

Abstract

Alternative splicing is a key layer of gene regulation that is frequently modulated in a spatiotemporal manner. As such, it is a major goal to understand the mechanisms controlling alternative splicing in specific cellular contexts. Reporters that recapitulate alternative splicing patterns of endogenous transcripts have served as excellent tools for dissecting regulatory mechanisms of splicing. In this chapter, we describe a two-color fluorescent reporter system that enables the visualization of alternative splicing patterns by microscopy at single-cell resolution in live animals. We present this reporter system in the context of the model nematode C. elegans.

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Acknowledgments

We would like to thank Dovic King and Lisa-Marie Schneider for their help with developing our updated two-color splicing reporters. Our research is currently supported by the Natural Sciences and Engineering Research Council of Canada (NSERC), the Canadian Institutes of Health Research (CIHR), the Canada First Research Excellence Fund (CFREF), the Canada Research Chairs program (CRC), the Canada Foundation for Innovation (CFI), and the Ontario Research Foundation (ORF) and we are grateful for their generous support.

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Authors and Affiliations

  1. Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada

    John A. Calarco & Pallavi P. Pilaka-Akella

Authors
  1. John A. Calarco
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  2. Pallavi P. Pilaka-Akella
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Corresponding author

Correspondence to John A. Calarco .

Editor information

Editors and Affiliations

  1. Biozentrum, University of Basel, Basel, Switzerland

    Peter Scheiffele

  2. Biozentrum, University of Basel, Basel, Switzerland

    Oriane Mauger

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Calarco, J.A., Pilaka-Akella, P.P. (2022). Two-Color Fluorescent Reporters for Analysis of Alternative Splicing. In: Scheiffele, P., Mauger, O. (eds) Alternative Splicing. Methods in Molecular Biology, vol 2537. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2521-7_13

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  • DOI: https://doi.org/10.1007/978-1-0716-2521-7_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2520-0

  • Online ISBN: 978-1-0716-2521-7

  • eBook Packages: Springer Protocols

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