Team:Freiburg Bioware/Project/Project Description
From 2010.igem.org
The Experimental System
Therapy using viral vectors is an promising approach for....
In an first step the plasmids of the AAV-2 Helper-free System were genetically modifyed by converting it into BioBricks and inserting of targeting molecules into the constructs. These plasmids were then used to transfect the producer cell line AAV-293. After an incubation of three days the viral vectors were harvested and used to transduce different target cells. The succesful transduction can then for example be measured by detecting the fluorescence of fluorescent proteins in the target cells.
The majority of the modifications that were introduced into the viral vector aimed to allow differential targeting of tumor cell over healthy off-target cells.
![](https://static.igem.org/mediawiki/2010/3/38/Freiburg10_Experimental_System.png)
The Experimental System
Employment of viral vectors for means of therapy is idea in the context of personalized medicie that gets more and more interest. In such applications the reduction of side effects and the safety of the patient in general is of the highest priority.
In order to satisfy this requirement we designed our Therapy Vector with several layers of Specificity:
![](http://partsregistry.org/wiki/images/b/b5/Freiburg10_Layers_of_Specificity.png)
Modularization of the rAAV genome
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Modulatization of the RepCap plasmid
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Modulatization of the Vectorplasmid
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![](https://static.igem.org/mediawiki/2010/8/8c/Freiburg10_Vectorplasmid.png)
Modification of the viral surface in one cloning step - The ViralBrick standard
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![](https://static.igem.org/mediawiki/2010/1/1b/Freiburg10_ViralBrick.png)
Midifications of the Virus Shell via Loop Insertions
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Specific biotinylation of the Viral Schell - The Biotinylation Acceptor Peptide (BAP)
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Purification of Therapeutic Viral Vectors - The His-Affinity Tag
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Targeting of integrin overexpressing cells - The RGD Motif
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Arming the Viral Vector with therapeutic antibodies - The Z34C Motif
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Testing the limit for loop insertion - The Beta-Lactamase
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Method Development
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Arming: Killing the Tumor
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All-In-One: Testing multiple modified Viral Vectors
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Virus Construction Kit - The Manual
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![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 ViralBrick.png](/wiki/images/1/1b/Freiburg10_ViralBrick.png)
![Freiburg10 Experimental System.png](/wiki/images/3/38/Freiburg10_Experimental_System.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)
![Freiburg10 Vectorplasmid.png](/wiki/images/8/8c/Freiburg10_Vectorplasmid.png)