Mar 12, 2016

Design of RNA-Binding Proteins: Manipulate Alternative Splicing in Human Cells with Artificial Splicing Factors

Methods in Molecular Biology
Yang Wang, Zefeng Wang

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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Mentioned in this Paper

Nested Transcripts
Subfamily lentivirinae
Shuttle Vectors
Transfection
Apoptosis, Intrinsic Pathway
Genes
Regulation of Biological Process
Splice Variants, Protein
RNA Splicing Factors
Human gene

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