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A robust and tunable mitotic oscillator in artificial cells

By Ye Guan, Zhengda Li, Shiyuan Wang, Patrick M. Barnes, Xuwen Liu, Haotian Xu, Minjun Jin, Allen P. Liu, Qiong Yang

Posted 16 Nov 2017
bioRxiv DOI: 10.1101/218263 (published DOI: 10.7554/elife.33549)

Single-cell analysis is pivotal to deciphering complex phenomena like cellular heterogeneity, bistable switch, and oscillations, where a population ensemble cannot represent the individual behaviors. Bulk cell-free systems, despite having unique advantages of manipulation and characterization of biochemical networks, lack the essential single-cell information to understand a class of out-of-steady-state dynamics including cell cycles. Here we develop a novel artificial single-cell system by encapsulating Xenopus egg extracts in water-in-oil microemulsions to study mitotic dynamics. These "cells", adjustable in sizes and periods, sustain oscillations for over 30 cycles, and function in forms from the simplest cytoplasmic-only to the more complicated ones involving nuclei dynamics, mimicking real mitotic cells. Such innate flexibility and robustness make it key to studying clock properties of tunability and stochasticity. Our result also highlights energy supply as an important regulator of cell cycles. We demonstrate a simple, powerful, and likely generalizable strategy of integrating strengths of single-cell approaches into conventional in vitro systems to study complex clock functions.

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