Abstract
The majority of human genes undergo alternative splicing to produce multiple isoforms with distinct functions. The dysregulations of alternative splicing have been found to be closely associated with various human diseases; thus new approaches to modulate disease-associated splicing events will provide great therapeutic potentials. Here we report protocols for constructing novel artificial splicing factors that can be designed to specifically modulate alternative splicing of target genes. By following the method outlined in this protocol, it is possible to design and generate artificial splicing factors with diverse activities in regulating different types of alternative splicing. The artificial splicing factors can be used to change splicing of either minigenes or endogenous genes in cultured human cells, providing a new strategy to study the regulation of alternative splicing and function of alternatively spliced products.
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Acknowledgements
This work is supported by NIH grant R01-CA158283 and the Jefferson Pilot award to Z.W. Y.W. is funded by the Young Thousand Talents Program and the National Natural Science Foundation of China (grants 31471235 and 81422038).
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Wang, Y., Wang, Z. (2016). Design of RNA-Binding Proteins: Manipulate Alternative Splicing in Human Cells with Artificial Splicing Factors. In: Lin, RJ. (eds) RNA-Protein Complexes and Interactions. Methods in Molecular Biology, vol 1421. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3591-8_18
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DOI: https://doi.org/10.1007/978-1-4939-3591-8_18
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3589-5
Online ISBN: 978-1-4939-3591-8
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