| Chapter title |
Use of the pBUTR Reporter System for Scalable Analysis of 3' UTR-Mediated Gene Regulation.
|
|---|---|
| Chapter number | 7 |
| Book title |
Post-Transcriptional Gene Regulation
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3067-8_7 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3066-1, 978-1-4939-3067-8
|
| Authors |
Arindam Chaudhury, Joel R. Neilson |
| Editors |
Erik Dassi |
| 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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 5 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Doctoral Student | 2 | 40% |
| Researcher | 1 | 20% |
| Student > Ph. D. Student | 1 | 20% |
| Unknown | 1 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 2 | 40% |
| Agricultural and Biological Sciences | 1 | 20% |
| Neuroscience | 1 | 20% |
| Unknown | 1 | 20% |