Gene of interest
The Virus Construction Kit enables researchers to encapsidate virtually any given DNA sequence into AAV-2 particles. As one example from therapeutic focus of our project, prodrug-activating enzymes are provided within the kit for efficient tumor cell killing. Additionally included fluorescent proteins allow monitoring of transduced cells by fluorescence microscopy and flow cytometry. Transgene expression can be fine-tuned using promoters of different specificity and enhancer elements also provided.
RepVP123
AAV-2 genes essential for the production of viral particles in a specialized cell line were identified by literature search, isolated on genetic level and modified to meet the requirements of BioBrick assembly. For this purpose, a specialized variant of the iGEM default backbone was created, proven functional and submitted.

One aim of our research is on the one hand to knock down the natural tropism of the Adeno-associates virus particles and on the other hand to specifically target tumor cells. This is achieved by genetic engineering of the virus surface. For this purpose, two different strategies were developed, including Targeting via Loops or fusion to the N-terminus of the viral protein VP2.

The specifically targeted tumor cells were killed by prodrug activation approaches. Viral particles were charged with thymidine kinase and cytosine deaminase constructs to induce apoptosis in cancer cells upon delivery of ganciclovir or 5-Fluorocytosine, respectively.