Transcripts for Combined Synthetic MicroRNA and Gene Delivery

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MicroRNAs (miRNAs) are a class of short noncoding RNAs which are endogenously expressed in many organisms and regulate gene expression by binding to messenger RNA (mRNA). MicroRNAs are either produced from their independent transcription units in intergenic regions or lie in intragenic regions. Intragenic miRNAs and their host mRNAs are produced from the same transcript by the microprocessor and the spliceosome protein complex respectively. The details and exact timing of the processing events have implications for downstream RNA interference (RNAi) efficiency and mRNA stability. Here we engineer and study in mammalian cells a range of synthetic intragenic miRNAs co-expressed with their host genes. Furthermore, we study transcripts which carry the target of the miRNA, thereby emulating a common regulation mechanism. We perform fluorescence microscopy and flow cytometry to characterize the engineered transcripts and investigate the properties of the underlying biological processes. Our results shed additional light on miRNA and pre-mRNA processing but importantly provide insight into engineering transcripts customized for combined delivery and use in synthetic gene circuits.;

MicroRNAs, Spliceosomes, Plasmids, Flow Cytometry, DsRed
"This work was funded by the US National Institutes of Health (NIH) grant GM096271, the US National Science Foundation (NSF) grant CBNET-1105524, and the University of Texas at Dallas."
©2013 The Royal Society of Chemistry
Kashyap, Neha, Bich Pham, Zhen Xie, and Leonidas Bleris. 2013. "Transcripts for combined synthetic microRNA and gene delivery." Molecular Biosystems 9(7): 1919-1925