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
Bleris L
中科院分区:
生物3区
文献类型:
--
作者:
Kashyap N;Pham B;Xie Z;Bleris L

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MicroRNAs (miRNAs)是一类短链非编码RNA,在许多生物体内内源性表达,并通过与信使RNA (mRNA)结合来调节基因表达。mirna要么由基因间区独立的转录单位产生,要么位于基因内区域。基因内mirna及其宿主mrna分别由微处理器和剪接体蛋白复合物从相同的转录物中产生。加工事件的细节和确切时间对下游RNA干扰(RNAi)效率和mRNA稳定性有影响。在这里,我们在哺乳动物细胞中设计和研究了一系列合成的与宿主基因共表达的基因内mirna。此外,我们研究了携带miRNA靶标的转录本,从而模拟了一种常见的调控机制。我们使用荧光显微镜和流式细胞术来表征工程转录本,并研究潜在生物过程的特性。我们的研究结果进一步揭示了miRNA和pre-mRNA的加工过程,但更重要的是,为在合成基因回路中联合传递和使用定制的工程转录本提供了见解。
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.
在含microRNA的内含子处的前馈微孔处理和剪接活性。
DOI: 10.1371/journal.pgen.1002330
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影响因子: 64.8
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