Collaborative Research: Transient Protein Expression in Plant Tissue Culture

The Principal Investigators (PIs)' overall objective is to develop plant cell and tissue culture technology, which will permit rapid evaluation of heterologous gene expression in plants. This work includes the development of rigorous principles for the monitoring, control and operation of bioreactors for plant cell and tissue culture. These bioreactor operational principles will be utilized to correlate performance of transient expression systems using Agrobacterium co-cultivation for DNA transformation and the TMV vector or a single-stranded DNA geminivirus vector capable of DNA amplification for protein production (based on tobacco and corn seed endosperm) while testing expression and accumulation of three therapeutic proteins: a monoclonal antibody (NR-LU-10 MAb), a cytokine (human growth hormone, hGH), and a blood transport protein (human transferrin, hTR). The specific objectives are to: (1) Develop operational and control strategies for bioreactor growth of plant cell and root culture, (2) Develop and optimize techniques / conditions for DNA delivery for transient expression, and (3) Utilize bioreactor control strategies and optimized transient therapeutic protein expression systems to evaluate heterologous transgene expression in plant cell and tissue culture, followed by correlation with performance in transgenic plants with the same proteins.

This is a collaborative GOALI project involving three PIs with different complementary scientific and technological expertise. Wayne Curtis the primary PI at the Pennsylvania State University is an expert in the area of bioreactor technology, Hugh Mason the second PI at the Boyce Thompson Institute and Cornell University in the area of transgenic plants, and Doug Russell at the Integrated Protein Technologies of Monsanto in the area of production of proteins from transgenic plants.