-Implementation of the system to synthesize and preserve a large amount of proteins in the identical cells-
NARO, in collaboration with RIKEN and Kazan Federal University (KFU, Russia) has discovered a novel promoter that allows synthesizing a large amount of protein in the dry-preservable insect cells. Moreover, this promoter can function in a wide variety of insect cells. This promoter could contribute to the effective production of practically useful proteins in various insect cultured cells.
The cultured cell line Pv11 established from the anhydrobiotic insect Polypedilum vanderplanki can be stored in a dry state at room temperature for as long as one year. Dried Pv11 cells allow the artificially synthesized proteins in the cells to be preserved for a long time with retaining their functions. However, the conventional promoters applicable for Pv11 cells enabled synthesis of an insufficient quantity of proteins in the cells.
A research group mainly consisting of NARO, RIKEN and KFU (Russia) has discovered a novel strong promoter in the process of the genome research of P. vanderplanki. This promoter designated as "121," showed approximately 1,500 times more capable of activity for gene expression than that of the promoters available in commercial kits for insect cells. By using the 121 promoter, Pv11 cells can now produce exogenous proteins massively. The 121 promoter also exerted strong promoter activity in other insect cells besides P. vanderplanki, such as the fruit fly and the silkworm.
This achievement must contribute to the development of effective production systems that combine "synthesis" and "preservation" of practically useful proteins.
Budget: This project was supported by the "Pilot program of international collaborative research (Collaborative research based on a joint call with Russia)" under "Commissioned projects for promotion of strategic international collaborative research".
1. Miyata Y., Tokumoto S., Sogame Y., Deviatiiarov R., Okada J., Cornette R., Gusev O, Shagimardanova E, Sakurai M & Kikawada T. (2019) Identification of a novel strong promoter from the anhydrobiotic midge, Polypedilum vanderplanki, with conserved function in various insect cell lines. Scientific Reports, 9(1), 7004. doi: 10.1038/s41598-019-43441-x
Fig 1. Gene expression activity of the 121 promoter in Pv11 cells
In Pv11 cells, relative gene expression activity of the 121 promoter was compared to that of the conventional Gapdh promoter (A) and the commercially-available IE2 promoter (B). The graph shows the means ± standard deviations of the activities. ***: p < 0.001.
Fig 2. The 121 Promoter can function in various insect cells