Part:BBa_K3743000

FAS-associated death domain protein (FADD)

Part description

FADD is an adaptor protein made up of 208 amino acids. It contains two main domains: a C terminal death domain (DD) and an N terminal death effector domain (DED). FADD forms the death-inducing signaling complex (DISC) during apoptosis when it interacts with other members of the tumor necrosis factor receptor superfamily, procaspases 8 and 10. Also, it has been shown to play a role in proliferation, cell cycle regulation, and development.

Usage

As our vaccine is supposed to be inserted intra-tumorally, we thought about using Fas-associated protein with death domain (FADD) to add an oncolytic feature to the circuit. FADD induces cancer cells apoptosis that combat TNBC cells in parallel with the vaccine activity. It will be linked to the vaccine to perform its function. FADD could be used by futuristic iGEM teams intending to establish immunotherapeutic approaches for cancer to add an oncolytic feature to their design.

Literature Characterization

A study tried to address the consequence of the lack of FADD and un-controlled T-cell necroptosis in an immune response setting, when infecting CD8-deficient micewith t.gondii they found out that they die within 20–25 d postinfection .they found that the absence of FADD renders mice more susceptible to chronic T. gondii infection. as shown also in figure (1)

Figure 1. Functional characterization of FADD from literature. This figure shows the effect of FADD as a negative regulator of t-cell specific necrosis & apoptosis compared to control(1)

Characterization Of Mutational Landscape

A mutational landscape prediction through saturation mutagenesis of FADD protein and the effect of these mutations on the evolutionary fitness of the protein is tested after generating multiple sequence alignment of the protein sequence and predict mutational landscapes. As shown in the chart, the (G58E) mutation showed the highest score compared to other mutations. On the contrary, we can see that the (V180E) contributed to the lowest evolutionary fitness to FADD. As shown in Figure (2)

Figure 2.shows the positive fit mutants upon saturation mutagenesis prediction of mutational landscape of FADD

References

1.Osborn, S. L., Diehl, G., Han, S. J., Xue, L., Kurd, N., Hsieh, K., ... & Winoto, A. (2010). Fas-associated death domain (FADD) is a negative regulator of T-cell receptor–mediated necroptosis. Proceedings of the National Academy of Sciences, 107(29), 13034-13039. Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]