Difference between revisions of "Part:BBa K4244000"

Revision as of 09:49, 4 October 2022

ALPaGA Lactate inducible promoter

For the proper functioning of our biosensor construct we must consider that it needs to function in a colonic environment. The lldPRD promoter does not function in the presence of glucose or the absence of oxygen. Unfortunately, both conditions are found in the human colon, meaning that PlldPRD¬ cannot be used to sense lactate in a colonic environment (Schwerdtfeger et al., 2019). Zúñiga et al. developed a lactate sensitive promoter called ALPaGA (A Lactate Promoter Operating in Glucose and Anoxia) (Zúñiga et al., 2021). This promoter is sensitive to lactate in the presence of glucose and absence of oxygen, perfect for developing a biosensor to sense colon cancer. Because we want to use ALPaGA to sense lactate produced by cancer cells, we set out to characterize and compare the native lldPRD operon promoter to ALPaGA. This was done by analysing both promoters in two different conditions: 1) absence of glucose, presence of oxygen and 2) presence of glucose, absence of oxygen, the results of which can be seen in Figure 1 and 2.

Sequence and Features

References

Reinfeld, et al.,(2021). Cell-programmed nutrient partitioning in the tumour microenvironment. Nature, 593(7858), 282–288. https://doi.org/10.1038/s41586-021-03442-1

Schwerdtfeger, L. A., Nealon, N. J., Ryan, E. P., & Tobet, S. A. (2019). Human colon function ex vivo: Dependence on oxygen and sensitivity to antibiotic. PLOS ONE, 14(5), e0217170. https://doi.org/10.1371/JOURNAL.PONE.0217170

Zúñiga, A., Camacho, M., Chang, H. J., Fristot, E., Mayonove, P., Hani, E. H., & Bonnet, J. (2021). Engineered l-Lactate Responding Promoter System Operating in Glucose-Rich and Anoxic Environments. ACS Synthetic Biology, 10(12), 3527–3536. https://doi.org/10.1021/acssynbio.1c00456