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Giant unilamellar vesicles (GUVs) provide a simplified analog of a cell membrane and can be utilized as minimal cell models for studying cellular systems due to their cell-like sizes and capacity to mediate membrane interactions. The OSM-PAPYRUS technique is a paper-based diffusive loading method that can be used to assemble GUVs in physiologically relevant salt solutions and allow the gentle loading of proteins. Characterization of this technique shows the encapsulated protein concentrations in the GUVs exhibit a cell-like variation, with a gamma distribution that is often observed for protein distributions in cells. By mimicking cellular variability, size, and membrane in vitro, GUVs can offer a more accurate representation of cellular systems than traditional in vitro studies and a simpler and more controlled environment than in vivo studies. The utility of GUVs is demonstrated by encapsulating the post-translation oscillator (PTO) of the cyanobacteria circadian clock system. This study showed that cellular variation and membrane binding significantly hampers the fidelity of the clock, highlighting the importance of other cellular components, such as SasA and CikA or the transcriptional-translational feedback loop (TTFL), in achieving in vivo clock fidelity. The well-characterized parameters of the GUV minimal cell model allow computation modeling to corroborate the effects of cellular variation and membrane binding on the behavior of the circadian clock. The ability of GUVs to mimic cellular variability and other cellular properties in vitro provides a promising avenue for bridging the gap between in vitro and in vivo experimentation and could facilitate the development of more accurate and predictive models of cellular systems. 



Alexander Li is a PhD candidate in Bioengineering at the University of California (UC), Merced, where he received his B.S. in Mechanical Engineering. Prior to joining Prof. Anand Subramaniam's Lab in 2018, he worked as a Junior Specialist in the UC Merced Bioengineering department under Prof. Subramaniam's mentorship. Li's research has focused on the use of giant lipid vesicles as minimal cell models for studying cellular systems, including the characterization of protein encapsulation in giant lipid vesicles and the study of the cyanobacteria circadian clock in these minimal cell models. His work has demonstrated the potential of giant lipid vesicles to bridge the gap between in vitro and in vivo experimentation.

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