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| - | {| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="70%" align="center" | + | {{Tsinghua/ProjectHeader}} |
| - | !align="center"|[[Team:Tsinghua/Project Original|Project Original]]
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| - | !align="center"|[[Team:Tsinghua/Background|Background]]
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| - | !align="center"|[[Team:Tsinghua/Brainsotrming|Brainsotrming]]
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| - | !align="center"|[[Team:Tsinghua/Design|Design]]
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| - | !align="center"|[[Team:Tsinghua/Experiment|Experiment]]
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| - | !align="center"|[[Team:Tsinghua/Results|Results]]
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| - | !align="center"|[[Team:Tsinghua/Notebook|Notebook]]
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| - | |}
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| | | | |
| - | = Project 1 =
| + | [[Image:project.png|500px|center|project.png|Overall Project|thumb|Three modules in our project]] |
| - | ==Synthesis of the Therapeutic DNA==
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| - | In this part, we aims at constructing a molecular cloning vector with a cos site which enables it to be packaged into the gene therapy vector- in other words, to construct a cosmid. In order to detect the expression of the therapeutic DNA, we construct this cosmid based on the scanfold of Parts-J61031 encoding a RFP-expressing segment.
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| - | This cosmid mainly consists of the following segments:
| + | Based on our basic idea and decoupling principle of synthetic biology, we subdivided our project into three interconnected modules: 1) Synthesis of the GenSniper virion which involves the synthesis of the GenSniper genome and the production of the viroin; 2) Constructing the Therapeutic DNA which can be packaged into the GenSniper virion; 3) Construction and characterization of the targeted BioBrick that is responsible for targeted gene delivery. The following wetlab work will demonstrate the synthesis of the GenSniper via the three modules respectively as well as how they work together. |
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| - | 1) origin of replication: we insert O gene and P gene from bacteriophage lambda into J61031, which are resoonsible for the late phase replication and package of the wild type circular bacteriophage lambda genome
| + | *[https://2009.igem.org/Team:Tsinghua/Experiment1 Experiment of ModuleI] |
| - | | + | *[https://2009.igem.org/Team:Tsinghua/Experiment2 Experiment of ModuleII] |
| - | 2) cos site: necessary for the specific package of the Therapeutic DNA into the gene therapy vector.
| + | *[https://2009.igem.org/Team:Tsinghua/Experiment3 Experiment of ModuleIII] |
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| - | 3) RFP-expressing segment: originally integrated into the plamid of J61031.
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| - | [[Image:cosmidcostruct.jpg|cosmidcostruct]] | + | |
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| - | ==Synthesis of the Gene Therapy Production System==
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| - | ===Bottom-Up Approach===
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| - | In the bottom-up approach, we amplify the biobricks from both the bacteriophage lambda and the adenovirus genome and incorporate them with a given order into molecular cloning vector(s).
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| - | Specifically, we choose two molecular cloning vectors to encode two sections of the gene therapy vector genome, pET28a and pACYCDuet1.
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| - | | + | |
| - | [[Image:pET28a.jpg|pET28a]]
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| - | [[Image:pACYCDuet1.jpg|pACYCDuet1]] | + | |
| - | | + | |
| - | ===Top-Down Approach===
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| - | | + | |
| - | ==Synthesis of the Targeted Biobrick==
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| - | ==Production of the targeted gene therapy vector==
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| - | ==Functioning of the targeted gene therapy vector==
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| - | | + | |
| - | = Project 2 =
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| - | ==Plasmid Construction==
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| - | Firstly, we build up a recombinant plasmid of pET-28a, into which a DNA sequence of lambda repressor is inserted, and we try to make it express the protein.
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| - | ==Targeted Biobrick Modification and Expression==
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| - | Secondly, another of DNA sequence is inserted into the plasmid, which can recognize and bind to the lambda repressor with specificity.
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| - | | + | |
| - | ==Transfection Efficiency Detection==
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| - | Thirdly, we can add another report gene, RFP sequence into the plasmid, and see whether it can be expressed. If everything goes all right, we can put the whole system in a eukaryotic cell.
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| - | | + | |
| - | = Project 1 and Project 2 =
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Based on our basic idea and decoupling principle of synthetic biology, we subdivided our project into three interconnected modules: 1) Synthesis of the GenSniper virion which involves the synthesis of the GenSniper genome and the production of the viroin; 2) Constructing the Therapeutic DNA which can be packaged into the GenSniper virion; 3) Construction and characterization of the targeted BioBrick that is responsible for targeted gene delivery. The following wetlab work will demonstrate the synthesis of the GenSniper via the three modules respectively as well as how they work together.
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