A robust and tunable mitotic oscillator in artificial cells
Ye Guan,
Zhengda Li,
Shiyuan Wang,
Patrick M Barnes,
Xuwen Liu,
Haotian Xu,
Minjun Jin,
Allen P Liu,
Qiong Yang
Affiliations
Ye Guan
Department of Biophysics, University of Michigan, Ann Arbor, United States; Department of Chemistry, University of Michigan, Ann Arbor, United States
Zhengda Li
Department of Biophysics, University of Michigan, Ann Arbor, United States; Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, United States
Shiyuan Wang
Department of Biophysics, University of Michigan, Ann Arbor, United States
Patrick M Barnes
Department of Physics, University of Michigan, Ann Arbor, United States
Xuwen Liu
Department of Physics, University of Science and Technology of China, Hefei Shi, China
Haotian Xu
Department of Computer Science, Wayne State University, Detroit, United States
Minjun Jin
Department of Biological Chemistry, University of Michigan, Ann Arbor, United States
Department of Biophysics, University of Michigan, Ann Arbor, United States; Department of Mechanical Engineering, University of Michigan, Ann Arbor, United States
Department of Biophysics, University of Michigan, Ann Arbor, United States; Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, United States; Department of Physics, University of Michigan, Ann Arbor, United States
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.
