Knockdown of Chimeric RNA by RNAi

Knockdown assays are widely used to study the functions of a gene of interest. RNA interference (RNAi) describes a set of well-known methods used to reduce the expression of a target gene by degrading its mRNA with short hairpin RNAs (shRNAs) or short interfering RNAs (siRNAs). Knockdown of chimeric...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2079; p. 143
Main Authors Qin, Fujun, Shi, Xinrui, Li, Hui
Format Journal Article
LanguageEnglish
Published United States 2020
Subjects
Cell Line
Gene Fusion
Gene Knockdown Techniques
Genetic Vectors
Humans
Real-Time Polymerase Chain Reaction
RNA - genetics
RNA Interference
RNA, Small Interfering - genetics
RNAi
Small interfering RNA (siRNA)
Chimeric RNA
Functional assay
Short hairpin RNA (shRNA)
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Summary:Knockdown assays are widely used to study the functions of a gene of interest. RNA interference (RNAi) describes a set of well-known methods used to reduce the expression of a target gene by degrading its mRNA with short hairpin RNAs (shRNAs) or short interfering RNAs (siRNAs). Knockdown of chimeric RNAs present different challenges than standard RNAi targeting for regular genes. Most specifically, sequence homology restricts the targeting region to the chimeric junction and can result in off-target effects on the parental genes. In this chapter, we provide guidelines and procedures for RNAi design of chimeric RNAs, knockdown of chimeric RNAs, downstream experiments for chimeric RNA functional studies and necessary controls to accompany each set of experiments.
ISSN:1940-6029
DOI:10.1007/978-1-4939-9904-0_11