Team:SUSTC-Shenzhen/Project/A-B toxin

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Novel DNA carrier protein TEG mimics the structure of Pseudomonas exotoxin A(ETA).
Novel DNA carrier protein TEG mimics the structure of Pseudomonas exotoxin A(ETA).
TEG acts as a vector transferring specific target DNA into cells via receptor-mediated endocytosis.   
TEG acts as a vector transferring specific target DNA into cells via receptor-mediated endocytosis.   
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ETA falls into three main structural and functional domains: the N terminal receptor (R) binding domain I, translocation (T) domain II and the catalytic (C) domain III. Here is the 3D structure of ETA.  The N terminal receptor binding domain is replaced by a human EGF receptor ligand TGF-a. EGF receptor is overexpressed by human carcinomas cells.  TEG containing TGF-a recognizes the EGF receptor and then binds to the EGF receptor. As a result, cells overexpressing EGF receptor are specifically combined with TEG. Similarly, replacing the C-terminal enzymatic domain of the toxin with the DNA-binding domain of the yeast GAL4 transcription factor leads to interaction with plasmid DNA of TEG. Overall, TEG contains TGF-a at N terminal following by translocation domain of ETA and GAL4 at C terminal
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ETA falls into three main structural and functional domains: the N terminal receptor (R) binding domain I, translocation (T) domain II and the catalytic (C) domain III. Here is the 3D structure of ETA.  The N terminal receptor binding domain is replaced by a human EGF receptor ligand TGF-a. EGF receptor is overexpressed by human carcinomas cells.  TEG(epidermal growth factor) containing TGF-a recognizes the EGF receptor and then binds to the EGF receptor. As a result, cells overexpressing EGF receptor are specifically combined with TEG. Similarly, replacing the C-terminal enzymatic domain of the toxin with the DNA-binding domain of the yeast GAL4 transcription factor leads to interaction with plasmid DNA of TEG. Overall, TEG contains TGF-a at N terminal following by translocation domain of ETA and GAL4 at C terminal
[[File:Structure_of_ETA.png‎ |frame|Fig. 1 Structure of ETA]]
[[File:Structure_of_ETA.png‎ |frame|Fig. 1 Structure of ETA]]
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===GD5===
===GD5===

Revision as of 20:18, 17 October 2014

Team SUSTC-Shenzhen

A-B Toxin

Adapted to Modular DNA Carrier Protein Work as Nucleic Acid Transfer System

Contents




Background

Though therapeutic techniques are developing rapidly nowadays, some human diseases such as cancer and AIDS are extremely difficult to effect a radical cure. Gene therapy cures diseases by using DNA that encodes a functional, therapeutic gene to replace a mutated gene. Therefore, gene therapy is efficient to cure diseases results from gene mutation such as cancer and chronic infectious diseases.

Two major methods are applied to gene therapy. One is viral vectors. The other is non-viral vectors. Viral vectors are efficient to transfer foreign gene into cells and gene is efficiently expressed. But, on the other hand, viral vectors are hard to target specific cells which will decrease the percentage of the target gene into target cells. Moreover, we can't sure that viral vectors are not infectious. It ceases to be a safe way for gene therapy. As a consequence, non-viral vectors gain increasing attention since the late 20th century. Several methods for gene therapy with non-viral vector are studied by researchers such as electroporation, gene gun, magnetofection and so on.

In our project, the novel modular DNA carrier protein is selected as one of the gene transfer vector. This nucleic acid transfer system was developed by Prof. Dr Wels from the Institute for Experimental Cancer Research, Tumor Biology Center and his cooperators in 1998.


Introduction

Our novel modular carrier protein is made up of three domains: Cell Binding Domain-specific binding to target cell, Translocation Domain- facilitating gene delivery via receptor-mediated endocytosis, DNA Binding Domain-specific binding to DNA containing UAS. Therefore, it can act as non-viral nucleic acid transfer system by delivering the carried target DNA to recognized the target cell via receptor-mediated endocytosis.

TEG

Novel DNA carrier protein TEG mimics the structure of Pseudomonas exotoxin A(ETA). TEG acts as a vector transferring specific target DNA into cells via receptor-mediated endocytosis. ETA falls into three main structural and functional domains: the N terminal receptor (R) binding domain I, translocation (T) domain II and the catalytic (C) domain III. Here is the 3D structure of ETA. The N terminal receptor binding domain is replaced by a human EGF receptor ligand TGF-a. EGF receptor is overexpressed by human carcinomas cells. TEG(epidermal growth factor) containing TGF-a recognizes the EGF receptor and then binds to the EGF receptor. As a result, cells overexpressing EGF receptor are specifically combined with TEG. Similarly, replacing the C-terminal enzymatic domain of the toxin with the DNA-binding domain of the yeast GAL4 transcription factor leads to interaction with plasmid DNA of TEG. Overall, TEG contains TGF-a at N terminal following by translocation domain of ETA and GAL4 at C terminal

Fig. 1 Structure of ETA

GD5

Mechanism

Purification

Transfer

Results

Prospects

Maintained by the iGEM team SUSTC-Shenzhen.

Licensed under CC BY 4.0.