Team:BAnner

From 2013.igem.org

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{{:Team:Korea_U_Seoul/template/banner}}
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<div style="width:100%; height:600px; background:white; padding:0 3px;">
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<title>Team:KAIST Korea/Project Background - 2013.igem.org</title>
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== '''Our Project Descriptions''' ==
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이 내용은 고대 생공 11학번 유민섭이 8월 9일까지 2문단 정도로 작성하기로 하였습니다.
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너무나 어렵군요! 뭘 더 써야할지 모르겟어yo....사실 자세한 내막을 잘몰라서..호호
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Our Team project is making pearl by E.coli induced Nacrein gene. At first, amplify kinds of nacrein genes by DH5a. Next, transformation of these genes to BW25113, BL21(DE3). Then nacrein genes make nacrein proteins. Finally they make nacrein aragonite structures itself and they forms the pearl.
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Nacrein has carbonic anhydrase domain that can make CO2 to carbonic acid, and it changes to bicarbonate ion(HCO3-) and H+. And We use Nacrein, Nacrein CA, Nacrein DR, Nacrein R gene to check what gene can make lots of pearl. (CA, DR and R indicate first carbonic anhydrase domain clone, repeat deleted-clone and only repeat clone, respectively.) We can make pearl with reducing CO2.
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The Korea_U_Seoul team aims to design Pearl-coli that is E. coli able to convert atmospheric CO2 into a pearl powder material. The design is based on cell surface display of nacrein in E. coli. Nacrein is a major protein component in nacre (an organic-inorganic composite layer found in outer coating of pearls). We divided nacrein into several functional regions - carbonic anhydrase (CA), calcium binding and scaffold repeats. Carbonic anhydrase domain fixes CO2 into carbonic acid changing to bicarbonate ion in aqueous solution. We assume that the other regions except CA is related to formation of nacre layer in a pearl.  The whole nacrein and those functional regions (CA, NR...??) are displayed in cell surface of E. coli. We will examine if displayed nacrein in E. coli can make a pearl powder in a solution or fabricate a nacre-like structure while atmospheric CO2 is fixed into bicarbonate. Once a nacre material can be prepared from Pearl-coli, we will grow E. coli in a confined container to make synthetic pearl. The Pearl-coli has dual-function such as (1) mitigate the global warming by CO2 reduction, (2) prepare valuable pearl-like raw materials.
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== 민환아 위키열렸다!!!!!!!!!!!!! 작업하자 ==
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<h1>Project : Overview</h1>
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<span id="little"><span id="starter">T</span>he widely used gene knock-out strategies are based upon a mere [on-off] control and more complex regulation is impossible. On the other hand, knock-down strategies are more versatile in usage but the current techniques are vastly laborious for systematic control and pose difficulty in multiple targeting because of the intricate design and scar accumulation.
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iGEM KAIST 2013 seeks to take the yet-to-be-perfected genetic regulation technology to a new level: The <span id="starter">RAPTOR</span> (RNA Associated Prokaryotic Transcript Optimization with CRISPR) technology. RAPTOR uses CRISPR type III system for gene regulation. CRISPR type III targets and cleaves specific mRNA sequences through complementary binding of crRNA and nuclease activity of Cmr protein complex. RAPTOR exploits this nature of CRISPR for systematically down regulating the mRNA level of specific genes.
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Due to the characteristics of RAPTOR, one of the most useful applications of this technology can be biosynthesis pathway regulation. Programmed crRNA expression allows fine-tuning of target mRNA expression levels and can precisely regulate enzyme concentrations. Not only fine-tuning but multiplex engineering is also possible through simultaneous expression of different crRNAs. Thus, multiple enzymes can be targeted in pathway control.
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As a proof of concept we sought to regulate the lycopene pathway that is incorporated into the genome of Escherichia coli. The efflux of lycopene precursors is optimally reduced through lowering the mRNA levels of enzymes that synthesize non-lycopene end products. Through observation of increased lycopene yield in the E.coli, potentiality of RAPTOR technology can be validated.
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Revision as of 01:54, 23 September 2013

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Project : Overview



The widely used gene knock-out strategies are based upon a mere [on-off] control and more complex regulation is impossible. On the other hand, knock-down strategies are more versatile in usage but the current techniques are vastly laborious for systematic control and pose difficulty in multiple targeting because of the intricate design and scar accumulation. iGEM KAIST 2013 seeks to take the yet-to-be-perfected genetic regulation technology to a new level: The RAPTOR (RNA Associated Prokaryotic Transcript Optimization with CRISPR) technology. RAPTOR uses CRISPR type III system for gene regulation. CRISPR type III targets and cleaves specific mRNA sequences through complementary binding of crRNA and nuclease activity of Cmr protein complex. RAPTOR exploits this nature of CRISPR for systematically down regulating the mRNA level of specific genes. Due to the characteristics of RAPTOR, one of the most useful applications of this technology can be biosynthesis pathway regulation. Programmed crRNA expression allows fine-tuning of target mRNA expression levels and can precisely regulate enzyme concentrations. Not only fine-tuning but multiplex engineering is also possible through simultaneous expression of different crRNAs. Thus, multiple enzymes can be targeted in pathway control. As a proof of concept we sought to regulate the lycopene pathway that is incorporated into the genome of Escherichia coli. The efflux of lycopene precursors is optimally reduced through lowering the mRNA levels of enzymes that synthesize non-lycopene end products. Through observation of increased lycopene yield in the E.coli, potentiality of RAPTOR technology can be validated.




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