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Issue 7, 2013
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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.

Graphical abstract: Transcripts for combined synthetic microRNA and gene delivery

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Publication details

The article was received on 30 Jan 2013, accepted on 22 Mar 2013 and first published on 26 Mar 2013

Article type: Paper
DOI: 10.1039/C3MB70043G
Citation: Mol. BioSyst., 2013,9, 1919-1925
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    Transcripts for combined synthetic microRNA and gene delivery

    N. Kashyap, B. Pham, Z. Xie and L. Bleris, Mol. BioSyst., 2013, 9, 1919
    DOI: 10.1039/C3MB70043G

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