Abstract
Posttranscriptional control of mRNA subcellular localization, stability, and translation is a central aspect of gene regulation and expression. Much of this control is mediated via recognition of a given mRNA transcript’s 3′ untranslated region (UTR) by microRNAs and RNA-binding proteins. Here we describe how a novel, scalable piggyBac-based vector, pBUTR, can be utilized for analysis of 3′ UTR-mediated posttranscriptional gene regulation (PTGR) both in vitro and in vivo. This vector is specifically designed to express a selection marker, a control reporter, and an experimental reporter from three independent transcription units. Expression of spliced reporter transcripts from medium-copy non-viral promoter elements circumvents several potential confounding factors associated with saturation and stability, while stable integration of these reporter and selection elements in the context of a DNA transposon facilitates experimental reproducibility.
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Chaudhury, A., Neilson, J.R. (2016). Use of the pBUTR Reporter System for Scalable Analysis of 3′ UTR-Mediated Gene Regulation. In: Dassi, E. (eds) Post-Transcriptional Gene Regulation. Methods in Molecular Biology, vol 1358. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3067-8_7
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DOI: https://doi.org/10.1007/978-1-4939-3067-8_7
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3066-1
Online ISBN: 978-1-4939-3067-8
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