Engineering Replication-, Growth-, and Division-Deficient E. coli for Safe, Stable and Efficient Function
The ultimate goal of this project is the ability to easily generate large quantities of E. coli cells with a durable capacity for performing engineered functions, but lacking the ability to replicate DNA, grow, and divide. This capability would not only prevent loss of engineered function due to mutation and natural selection or genetic drift, but also directly address concerns of escape of engineered cells into the environment, as well as improve efficiency of bioproduction by reducing the portion of feedstock incorporated into cell biomass. As we pursue this goal, we will be forced to interrogate the impact of preventing genome replication/cell growth and division on E. coli physiology, metabolism, and gene expression, in order to intervene appropriately to adapt cells to this condition. This may not only generate new knowledge of E. coli physiology and gene regulation, but also inform how cells may be adapted to perform well in the context of engineered living materials, where spatial constraints necessarily limit cell growth and division.
In this project we will demonstrate the ability to create "simple cells" that can serve as a non-living chassis for carrying out engineered bimolecular functions. Our specific plans for the project are:
- Generating pure populations of DNA replication-deficient cells
- Improving viability and lifespan of DNA replication-deficient cells
- Improving capacity of DNA replication-deficient cells for engineered functions
None to date
This research was supported by the Rosen Center for Bioengineering