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A robust and tunable mitotic oscillator in artificial cells.

eLife (2018-04-06)
Ye Guan, Zhengda Li, Shiyuan Wang, Patrick M Barnes, Xuwen Liu, Haotian Xu, Minjun Jin, Allen P Liu, Qiong Yang
ABSTRACT

Single-cell analysis is pivotal to deciphering complex phenomena like heterogeneity, bistability, and asynchronous oscillations, where a population ensemble cannot represent 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, by encapsulating Xenopus egg extracts in water-in-oil microemulsions, we developed artificial cells that are adjustable in sizes and periods, sustain mitotic oscillations for over 30 cycles, and function in forms from the simplest cytoplasmic-only to the more complicated ones involving nuclear dynamics, mimicking real cells. Such innate flexibility and robustness make it key to studying clock properties like tunability and stochasticity. Our results also highlight energy 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.

MATERIALS
Product Number
Brand
Product Description

Glutathione Sepharose 4B, Cytiva 17-0756-01, pack of 10 mL
Disposable PD 10 Desalting Columns, Cytiva 17-0851-01, pack of 30 pieces
Sigma-Aldrich
Trichloro(1H,1H,2H,2H-perfluorooctyl)silane, 97%