Development of an inducible single lentiviral vector-based microRNA knockdown construct.
MicroRNAs (miRNAs) have emerged as important post-transcriptional developmental regulators of gene expression in normal cellular processes. Perturbations in the levels of individual miRNAs or classes of miRNAs are associated with the pathogenesis of a wide range of human diseases. RNA interference (...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2013
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| Online Access: | http://hdl.handle.net/10356/52927 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | MicroRNAs (miRNAs) have emerged as important post-transcriptional developmental regulators of gene expression in normal cellular processes. Perturbations in the levels of individual miRNAs or classes of miRNAs are associated with the pathogenesis of a wide range of human diseases. RNA interference (RNAi) is an efficient approach for rapid analysis of gene functions. The current limitation in studying the biological functions of miRNAs lie in the lack of an inducible system that conditionally knockdown miRNA expression. With some modifications of a previously described inducible system that repress gene expression, we developed a lentivector-based, conditional gene expression system for drug-controllable expression of antimiRs, using miR-138 as our model of study. We present evidence that antimiRs can be transcribed from RNA polymerase III-driven H1 promoter with seven upstream tetO sequences in a tTRKRAB-controlled manner, permitting the knockdown of endogenous miR-138. This methodology should expedite large-scale functional studies. A better understanding of the mechanisms underlying miRNAs could provide new insights into the pathogenesis of diseases and open up new perspectives for development of therapeutic modalities. |
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