Issue 7, 2013
From the journal:

Molecular BioSystems

Transcripts for combined synthetic microRNA and gene delivery†

Neha Kashyap,ab   Bich Pham,c   Zhen Xie*d  and  Leonidas Bleris*abe  

* Corresponding authors

a Bioengineering Department, The University of Texas at Dallas, 800 West Campbell Road, Richardson, TX 75080, USA
E-mail: bleris@utdallas.edu

b Center for Systems Biology, The University of Texas at Dallas, NSERL 4.708, 800 West Campbell Road, Richardson, TX 75080, USA

c Bioengineering Department, The University of Texas at Arlington, 701 South Nedderman Drive, Arlington, TX 76019, USA

d Center for Synthetic and Systems Biology, Bioinformatics Division, TNLIST, Tsinghua University, Beijing, China
E-mail: zhenxie@tsinghua.edu.cn

e Electrical Engineering Department, The University of Texas at Dallas, 800 West Campbell Road, Richardson, TX 75080, USA

Abstract

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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Article information

DOI
https://doi.org/10.1039/C3MB70043G
Article type
Paper
Submitted
30 Jan 2013
Accepted
22 Mar 2013
First published
26 Mar 2013

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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