Team:Tsinghua/Project

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!align="center"|[[Team:Tsinghua|Home]]
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=Overall Project=
 
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A relatively significant procedure in gene therapy is to construct a vector to infect target cells and deliver cure gene into them. As a result, the vectors act as big role. And till now researchers use Adeno-associated viruses to do a good job, but the problems of high cost and low production of the virus has not been solved. That is why we attempted to build a highly productive carrier into bacteria. We transformed the structure genes of the phage into bacteria with specific chimera genes attached to the structural genes. We attempted to simulate the Adeno-associated viruses by the phage, for they share the similar structure. Fiber has been attached to the phage to enhance transformation efficiency. Not only we use the cosmid in the phage to carry the cure gene, which has great capability to carry large and multiple genes, but also the cure genes are tissue specific.
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=Overall Project=
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Our aim is to construct a targeted gene therapy vector with high cellular specificity, considerable capacity and the potential for mass production and universal modification. Analogizing the characteristics of bacteriophage lambda and adenovirus, we genomically engineered the fiber protein of adenovirus with the pC of bacteriophage lambda, with the knob region modified by cell-specific peptides generated by phage display (called targeted bioBrick). After inducing the vector genome (generated by bottom-up or top-down approach) into BL21 DE3 E.coli strain, we applied a co-transformed therapeutic DNA (namely a cosmid with a capacity of 40-50 kb) for mass production of our targeted gene therapy vectors containing the desired genes to be delivered. With the targeted bioBrick mediating the attachment and RGD domain mediating the internalization of the targeted vector, we are able to accomplish the targeted gene therapy.
==ModuleI: Synthesis of GenSniper Viroin==
==ModuleI: Synthesis of GenSniper Viroin==
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Using two synthetic biological strageties (bottom-up and top-down), we will construct a genome of our desired gene therapy vector, which consists of the structural proteins for bacteriophage lambda head assembly and lysis genes (and targeted bioBrick later). Our aim in this module is to generate the viroin of our gene therapy vector (namely the head structure of bacteriophage lambda).
==ModuleII: Therapeutic DNA==
==ModuleII: Therapeutic DNA==
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In this module, we plan to construct a cosmid (a molecular cloning with a cos site) encoding the conceptual therapeutic genes, because the structural proteins of bacteriophage lambda will only package the circular DNA with cos site and 40-50 kb in size. Here we will also identify the function of this module through the expression of RFP.
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==ModuleIII: Targeted BioBrick==
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This module is the modification of bacteriophage lambda protein C by fiber shaft at its N termini. When constructing this targeted bioBrick, we will apply PCR to add the targeted peptide (in wetlab we just use TAT peptide) and RGD sequence into the fusion protein, forming a peptide-fiber shaft-RGD-C reading frame.
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==ModuleIII: Targeted Biobrick==
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{{Tsinghua/Pic}}

Latest revision as of 00:05, 22 October 2009

Home Background Brainstorming Design Experiment Results Conclusion Protocol


Overall Project


Contents

Overall Project

Our aim is to construct a targeted gene therapy vector with high cellular specificity, considerable capacity and the potential for mass production and universal modification. Analogizing the characteristics of bacteriophage lambda and adenovirus, we genomically engineered the fiber protein of adenovirus with the pC of bacteriophage lambda, with the knob region modified by cell-specific peptides generated by phage display (called targeted bioBrick). After inducing the vector genome (generated by bottom-up or top-down approach) into BL21 DE3 E.coli strain, we applied a co-transformed therapeutic DNA (namely a cosmid with a capacity of 40-50 kb) for mass production of our targeted gene therapy vectors containing the desired genes to be delivered. With the targeted bioBrick mediating the attachment and RGD domain mediating the internalization of the targeted vector, we are able to accomplish the targeted gene therapy.

ModuleI: Synthesis of GenSniper Viroin

Using two synthetic biological strageties (bottom-up and top-down), we will construct a genome of our desired gene therapy vector, which consists of the structural proteins for bacteriophage lambda head assembly and lysis genes (and targeted bioBrick later). Our aim in this module is to generate the viroin of our gene therapy vector (namely the head structure of bacteriophage lambda).

ModuleII: Therapeutic DNA

In this module, we plan to construct a cosmid (a molecular cloning with a cos site) encoding the conceptual therapeutic genes, because the structural proteins of bacteriophage lambda will only package the circular DNA with cos site and 40-50 kb in size. Here we will also identify the function of this module through the expression of RFP.

ModuleIII: Targeted BioBrick

This module is the modification of bacteriophage lambda protein C by fiber shaft at its N termini. When constructing this targeted bioBrick, we will apply PCR to add the targeted peptide (in wetlab we just use TAT peptide) and RGD sequence into the fusion protein, forming a peptide-fiber shaft-RGD-C reading frame.


Tsinghua06.jpeg