Transcripts for combined synthetic microRNA and gene delivery.
Transcripts for combined synthetic microRNA and gene delivery.
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DOI:
10.1039/c3mb70043g
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发表时间:
2013-07
影响因子:
--
通讯作者:
Bleris L
中科院分区:
文献类型:
--
作者:
Kashyap N;Pham B;Xie Z;Bleris L
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). MiRNAs 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 to 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 towards engineering transcripts customized for combined delivery and use in synthetic gene circuits.
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