The nucleus is where a cell stores information. The endoplasmic reticulum (ER) is where cells manufacture proteins. Mitochondria are where cells generate energy. Designing cells to mimic human cells will likely involve replicating at least those core functions. This project will attempt to do just that. The project will involve artificial chromosomes being packaged into a compartment surrounded by a membrane, representing the nucleus. The nucleus will be surrounded by a folded membrane structure that mimics the ER. Inside the ER will be machinery to produce proteins. A separate membrane compartment, unattached to the other two, will house enzymes for generating energy. All of these will be contained in another membrane compartment and together, represent a crude synthetic cell. Creating the individual compartments will be challenging because the membranes have to allow some molecules to pass through them selectively while preventing others, and because some membranes will need to have intricate shapes. If the project is successful, it will provide a platform for subsequent efforts to use these types of synthetic cells to manufacture vaccines or to kill tumors, just to name a couple of possibilities. The project will involve a team of local high school students in all phases of the research through the International Genetically Engineered Machine (iGEM) competition.
This project will develop the techniques needed to engineer cell-like structures, enabling the development of complex synthetic systems mimicking both structure and function of biological cells. There are two major tasks. First, microfluidic techniques will be employed to assemble a defined population of chromosomes within a discrete aqueous volume. A nuclear envelope will be formed around the chromosomes and interconnected with a folded ER-like membrane engineered using curvature-promoting proteins. Biological components extracted from cells will be integrated into the membrane. A nuclear lamina will be formed within the nucleus. Transcription of mRNA within the synthetic nuclei will then be demonstrated. Second, mitochondria-like structures will be developed to serve as ATP sources for mRNA transcription. A vesicle-based approach will support external control over mRNA transcription through mitochondria-nucleus ATP transport. This project will contribute to understanding the requirements for synthesizing and linking other cell-like organelles with increasing levels of complexity. That will create new opportunities for exploring open questions such as membrane how and why cells age. Synthetic cells could ultimately be exploited for closed loop control over DNA processing and gene regulation, with significant potential for practical biomedical applications including cancer immunotherapy.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
