원문정보
초록
영어
Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) fusion protein was produced from transgenic rice cell suspension cultures using RAmy3D promoter as a novel immunosuppressive agent. Since RAmy3D promoter was strongly induced by sugar starvation, both cell viability and protein productivity were reduced remarkably after induction. To overcome these obstacles, various strategies were contemplated to develop the production processes of hCTLA4Ig. The expression of proteases and repressors was reduced by siRNA. To apply siRNA technology into transgenic rice cell cultures, three steps should be optimized, to put it briefly, 3D steps: 1) Design of siRNA sequences, 2) Delivery of siRNA into rice cells, and 3) Detection of the inhibited level of target proteins. siRNA sequences were designed by web-based programs like siRNA Target Finder, siDESIGN Center, and BLOCK-iT RNAi Designer. siRNA was combined with polyethyleneimine (PEI) to form siRNA complexes for the delivery into plant cells. Moreover, sonoporation was practically treated to enhance the transport of siRNA complexes.