Team:Heidelberg/Project

From 2010.igem.org

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<div class="t1">Abstract</div><br>
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<div class="t3">miBricks: DNA is not enough</div>
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<div id="wrapperheadline">iGEM Heidelberg Mission 2010: miBricks</div><br><br>
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<div id="projectabstract">The key to successful gene therapy is integration of tissue specificity and fine-tuned target gene expression. iGEM Team Heidelberg 2010 unlocks the world of synthetic microRNAs. We engineered a toolkit for standardized measurements of interactions between artificial miRNAs and their binding sites. Thus, the expression level of any gene of choice could be arbitrarily adjusted by employing the corresponding binding site design. To produce tissue specific miRNA gene shuttles, we developed an evolution-based method for synthesis of new adeno associated viruses. In the future, miBricks could open the doors to new Synthetic Biology based medical approaches. </div>
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<div id="projectabstract">The iGEM Team Heidelberg 2010 unlocks the entirely new field of synthetic mi(cro)RNA technologies for mammalian cell and tissue engineering. By combining novel miRNA-based tools and protocols with cell-specific viral delivery systems our technology allows for the first time to use RNAi for rationally engineering gene regulation in targeted cells and organs. The two most prominent technologies we developed are a new measurement standard for real-time detection/quantification of miRNA activity in living cells (miMeasure) as well as a synthetic miRNA expression kit (miTuner) that allows to precisely repress (fully or partly) and thus fine-regulate any desired target gene. The rational design of this miRNA expression kit is accomplished by a novel computational model for miRNA-based gene silencing termed miBEAT. The miRNA technology is accompanied by a novel gene delivery technology based on re-designed Adeno-associated viruses (AAV) that were molecularly engineered and evolved to specifically deliver our miRNA expression kit to hepatocytes. The rationally designed synthetic miRNA expression kits were successfully validated in cultured, transformed or primary liver cells and then transferred into an adult mouse model. We thereby demonstrate that our miRNA expression kit is able to specifically fine tune the expression level of target genes both in cell culture (in vitro) and importantly also in the liver of mice (<i>in vivo</i>). In summary, we show that this technology allows the precise, predictable and quantitative adjustment of mammalian gene expression levels. Our work fosters the introduction of synthetic biology based technologies into the rapidly emerging field of personalized biomedicine.</div>
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<div style="font-weight:bold">The circle of gene regulation</div>
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<p>Fine-tuning of gene expression was achieved by application of miRNAs, both endo- and exogenous ones, as well as by the use of AAVs.</p>
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<img src="http://2010.igem.org/wiki/images/2/26/RegCircle.png"/>
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Revision as of 00:36, 28 October 2010

Abstract

miBricks: DNA is not enough
The iGEM Team Heidelberg 2010 unlocks the entirely new field of synthetic mi(cro)RNA technologies for mammalian cell and tissue engineering. By combining novel miRNA-based tools and protocols with cell-specific viral delivery systems our technology allows for the first time to use RNAi for rationally engineering gene regulation in targeted cells and organs. The two most prominent technologies we developed are a new measurement standard for real-time detection/quantification of miRNA activity in living cells (miMeasure) as well as a synthetic miRNA expression kit (miTuner) that allows to precisely repress (fully or partly) and thus fine-regulate any desired target gene. The rational design of this miRNA expression kit is accomplished by a novel computational model for miRNA-based gene silencing termed miBEAT. The miRNA technology is accompanied by a novel gene delivery technology based on re-designed Adeno-associated viruses (AAV) that were molecularly engineered and evolved to specifically deliver our miRNA expression kit to hepatocytes. The rationally designed synthetic miRNA expression kits were successfully validated in cultured, transformed or primary liver cells and then transferred into an adult mouse model. We thereby demonstrate that our miRNA expression kit is able to specifically fine tune the expression level of target genes both in cell culture (in vitro) and importantly also in the liver of mice (in vivo). In summary, we show that this technology allows the precise, predictable and quantitative adjustment of mammalian gene expression levels. Our work fosters the introduction of synthetic biology based technologies into the rapidly emerging field of personalized biomedicine.
The circle of gene regulation

Fine-tuning of gene expression was achieved by application of miRNAs, both endo- and exogenous ones, as well as by the use of AAVs.