Team:LMU-Munich/Apo Control

The control of gene expression in eukaryotic systems has been challenging due to the complexity of structure and physiology of eukaryotic cells. The project ApoControl is focused on developing a system to improve the efficiency and specificity of gene expression in eukaryotic cell-lines. Here proapoptotic gens instead of antibiotic resistance are used as selection marker to induce clean cell-death at different stimuli. The project is divided into three parts, each representing different approaches to achieve this goal.

1) the TEV-Degron System: This system utilizes the property of N-Degron to degrade the proapoptotic protein when target gene is successfully transfected into the cell. The recognition and activation of N-Degron is carried out by TEV-protease. Thus this system enables selection of cells expressing the target gene while the cells not transfected undergo induced apoptosis.

2) The Jump-and-Stop System: This system makes use of the PhiC31o integrase and corresponding recombining sequences to integrate a target gene into one specific site in the chromosome of the cell line. Cells with integrated target gene will not express the proapoptotic gene due to the stop codon after the target gene while other cells undergo induced apoptosis.

3) The ApoPredict System: This part is carried out by bioinformatic modeling process and the end-product is an interacting apoptosis-inducing/prohibiting network which would take the interaction between different apoptosis-related genes into consideration when predicting the outcome of certain stimuli. This system will be useful for the construction of new systems trying to utilize apoptosis in eukaryotic cells.