title: Transcripts for combined synthetic microRNA and gene delivery.
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.
authors.affiliation: Bioengineering Department, The University of Texas at Dallas, 800 West Campbell Road, Richardson, TX 75080, USA.
authors.forename: Neha
authors.initials: N
authors.lastname: Kashyap
authors.forename: Bich
authors.initials: B
authors.lastname: Pham
authors.forename: Zhen
authors.initials: Z
authors.lastname: Xie
authors.forename: Leonidas
authors.initials: L
authors.lastname: Bleris
mesh_terms: D002460:Cell Line; D015870:Gene Expression; D005786:Gene Expression Regulation; D023061:Gene Order; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012326:RNA Splicing; D014158:Transcription, Genetic
chemical_list: D035683:MicroRNAs