Part:BBa_K3743013

BAX mutant (G36E)

Part Description

This is a Bax mutant (G36E) improved from mutational landscape prediction through saturation mutagenesis that is a protein encoded by a gene, which is a member of the BCL2 family. It regulates a variety of cellular activities, including a role in the mitochondrial apoptotic process. Under normal circumstances, BAX is cytosolic due to constant retrotranslocation from the mitochondria to the cytosol mediated by BCL2L1/Bcl-xl, thereby avoiding accumulation at the mitochondrial outer membrane of toxic levels. However, under stress conditions, translocation to the mitochondrion membrane occurs, leading to the release of cytochrome c, which triggers apoptosis. So It acts as a key gene in our biosafety device.

Usage

Our TNBC vaccine is supposed to be injected intra-tumorally. That is why an oncolytic activity is added to the circuit to perform an apoptotic function which helps in both combating cancer cells and apoptotic bodies which release neo-epitopes to be recognized by immune cells.

Literature Characterization

A Study found out that HBAX is a strong suppressor of tumorigenesis as well as stimulating apoptosis in vivo. And in one experiment done on mice they found out that apoptosis inversely correlates with tumour growth rate in which different variants of BAX with 3 Different experimental sets of mice that aged 3.5 weeks, 4 weeks and 6 weeks respectively.as shown in figures (1) & (2). Also as shown in figure (3) they found out that Bax deficiency accelerates tumour growth indicated by the mean survival time of mice that were deficient in variants of BAX as well as inactive p53. growth, as indicated by a decrease in survival time

Figure 1.Percentage of apoptotic cells in mice of the 3 different cells using different variants of BAX

Figure 2.relative apoptosis rate in comparison to mean survival time indicative of survival time derived from figure (3)

Figure 3.percentage of tumor survivors from mice on the vertical axis as well as the age of survival per week in experimental sets that included different groups of mice with either deficiency of a variant of bax or p53

Characterization Of Mutational Landscape

After performing mutagenesis prediction of mutational landscape of Bax mutant and tested the effect of these mutations on the evolutionary fitness of the protein after generating multiple sequence alignment of the protein sequence and predict mutational landscapes. As shown in the chart, the (G36E) mutation showed the highest score compared to other mutations. On the contrary, we can see that the (G36K) contributed to the lowest evolutionary fitness to Bax mutant. As shown in Figure (4) While also characterizing & improving part BBa_K2365048

Figure 4.shows the positive fit mutants upon saturation mutagenesis prediction of mutational landscape of BAX mutant

References

1.Yin, C., Knudson, C. M., Korsmeyer, S. J., & Van Dyke, T. (1997). Bax suppresses tumorigenesis and stimulates apoptosis in vivo. Nature, 385(6617), 637-640.

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]