5. Process-regulated gene expression based on autologously regulated endogenous promoters

It has been shown that even thought there is no tight correlation between growth and productivity, typically very highly producing cell lines exhibit a slower growth compared to lower producers. Based on this, many processes use for instance temperature shift to reduce growth at defined time points and enhance productivity, while maintaining viability. The use of CHO autologous promoters that are integrated into the regulatory network of the cells and which drive expression at low levels under growth conditions, but are highly active during defined process conditions (fi. Temperature shift) would enhance the overall yield and help in speeding up the process. Such promoters can be identified from transcriptomics data, indeed a list of potential genes is already available. Such promoters could also be used for cell engineering, if the expression of regulatory proteins is desired in a controlled and cell state adapted way.

Approaches:

  • Identification and testing of the promoters of a short list of regulated or highly expressed genes
  • Experimental validation and characterisation of these promoters
  • General screen of the genomic sequence for promoters


Target: Short-list of CHO endogenous promoters and their characteristics