Overexpression of RasV12 and its role in promoting the robustness of HEK293T cells.

• Main Author: Chow, Peiyi Vicki.
• Other Authors: School of Biological Sciences
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Science::Biological sciences::Molecular biology
• Online Access: http://hdl.handle.net/10356/52745
• Institution: Nanyang Technological University

Mammalian cells are usually grown in stirred bioreactors for the production of biopharmaceuticals in which the proliferation and health of the cells are affected by the limiting factors such as environment and growth inhibition. To overcome such limitations, cell engineering, such as the overexpression of oncogenes, has been employed in recent years (Mohan et al., 2008). In this study, RasV12 was overexpressed in HEK 293T cells and grown in several conditions – complete growth medium, low glucose, glucose free, low serum, serum free and media supplemented with either lactate or ammonium over 6 days. It was observed that RasV12 expressing cells managed to obtain higher cell numbers and viability as compared to the controls generally in all conditions. Thus this indicates that the overexpression of RasV12 conferred resistance to adverse conditions. However for the glucose and serum conditions, no significant differences was detected using ANOVA in cell numbers and viability against the controls (p>0.05).These results denotes that cells expressing RasV12 are dependent on nutrients such as glucose and serum to drive proliferation. In conclusion, the transfection and overexpression of oncogenes such as RasV12 shows promise in the optimization of mammalian cells for the production of biopharmaceuticals.