Confirmation of Transcriptional Read-Through Events by RT-PCR

Chimeric RNAs can be formed by trans-splicing from different transcripts or cis-splicing of adjacent genes (cis-SAGe). Cis-SAGe results from read-through transcription of two neighbor genes. To investigate the mechanisms underlying intergenic splicing of adjacent genes, it is important to develop an...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2079; p. 177
Main Authors Shi, Xinrui, Qin, Fujun, Li, Hui
Format Journal Article
LanguageEnglish
Published United States 2020
Subjects
Genetic Loci
HeLa Cells
Humans
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction - methods
RNA Precursors
RNA Splicing
RNA, Messenger
Transcription, Genetic
Reverse transcription
cis-SAGe
Intergenic splicing
Precursor mRNA
PCR
Transcriptional read-through
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Summary:Chimeric RNAs can be formed by trans-splicing from different transcripts or cis-splicing of adjacent genes (cis-SAGe). Cis-SAGe results from read-through transcription of two neighbor genes. To investigate the mechanisms underlying intergenic splicing of adjacent genes, it is important to develop an assay to detect transcriptional read-through. Here, we describe a general RT-PCR based method to confirm the process for cis-SAGe candidates. In this method, we use PCR to amplify cDNA that is reverse transcribed from the read-through precursor mRNA. The result provides a foundation for further downstream mechanistic studies.
ISSN:1940-6029
DOI:10.1007/978-1-4939-9904-0_14