The manipulation of giant vesicles and their use as model cells
Open Access
- Author:
- Gundermann, Erica Lynn
- Area of Honors:
- Chemistry
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Christine Dolan Keating, Thesis Supervisor
Christine Dolan Keating, Thesis Supervisor
Raymond Lee Funk, Thesis Honors Advisor - Keywords:
- giant vesicles
model cells
synthetic cells
microcompartmentation
encapsulation efficiency - Abstract:
- Giant vesicles are often used as model cells. They allow for a simplified system that can be prepared and manipulated in a variety of ways. In this thesis, we focused on the manipulation of giant vesicles, their encapsulation of solutes and microcompartmentation. First, the encapsulation efficiency (EE) of a larger protein was greatly improved by using PEG as a crowding agent. Also, using an aqueous two-phase system (ATPS), microcompartments were created that mimic the compartments within a living cell. A protein was localized to one phase and relocalized to the other phase, by an external change in pH. Finally, we aimed to create vesicles that more closely modeled living cells, so that they may be used for biological research. For this purpose, we aimed to develop methods of vesicle formation that produced more abundant and homogeneous populations of vesicles. Methods of vesicle formation were improved with an adaptation of the gentle hydration method that used agarose as a pre-hydration step for vesiculation. Giant vesicles were also created in high ionic strength buffer in order to imitate the environment of living cells; however, the giant vesicles were often heterogeneous and did not form consistently. In the future, methods of formation may be further altered, so that giant vesicles can more accurately represent living cells.