Team:Freiburg/Collaborations

Overview Motivation CAR T Cells Tumor Microenvironment AND Gate Outlook Achievements References

Members Attributions Partners Contact

Main Project Modeling Applied Design Proof of Concept BioBricks Basic Part BioBrick Improvement Interlab Study

Safety Cloning HIF1A Knockdown Cell Culture Lab Book

Human Practice Integrated Human Practice Collaborations

Collaborations

Zurich

Transfection

Mammalian cell transfection is used to introduce foreign nucleic acids into cells. Depending on cell line and desired effect, there are different ways to transfect mammalian cells. These methods can be broadly classified into two groups: transient and stable transfections. Normally, a selection marker is needed to identify cells that are transfected.

Heidelberg

Under normoxic conditions HIF1A cannot interact with HIF1B to form the transcription factor HIF1, because its hydroxylated and marked by the E3 ubiquitin ligase for degradation by the proteasome (Ziello, J. et al., 2007).

Therefore the elimination of the two hydroxylated amino acids, that are prolines at the positions 402 and 564, would generate a HIF1A, that is stabilized and interacts with HIF1B under normoxic conditions (Yasui et al., 2017) to activate the hypoxia response element (HRE).

Because we wanted to collaborate with this year’s iGEM team Heidelberg , they offered to help us with their Golden Gate cloning expertise to perform a site directed mutagenesis and generate the constitutively active HIF1A. It would be used by us as a positive control for CoCl2 induction in analysis of HRE.

Integrated HP

Team