A robust and tunable mitotic oscillator in artificial cells

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.

Inhibition of PFKFB3 induces cell death and synergistically enhances chemosensitivity in endometrial cancer

“…Metabolic reprogramming has been widely accepted as a distinctive hallmark of cancer. Cancer cells exhibit increased glucose uptake, enhanced glycolysis and lactate production, and reduced oxygen consumption, even in oxygen-rich conditions, known as the Warburg effect or aerobic glycolysis [24][25][26]. In addition to promoting tumorigenesis, aerobic glycolysis plays a vital role in developing drug resistance [27].…”

Organelle size scaling over embryonic development

“…As already mentioned, reductions in cell size over early development are driven by rapid cell divisions without cell growth, so cell cycle timing is a key determinant of cell size in the early embryo. There is also evidence that cell size and cytoplasmic volume determine cell division timing, with cell size reductions leading to increased cell cycle lengths (Arata & Takagi, 2019;Guan et al, 2018;Masui & Wang, 1998). Characteristic of many early developmental programs, asymmetric cell divisions give rise to different sized cells within the embryo (Cadart, Zlotek-Zlotkiewicz, Le Berre, Piel, & Matthews, 2014).…”

Cell-free microcompartmentalised transcription–translation for the prototyping of synthetic communication networks

“…It has been established that size and confinement have a considerable impact on TXTL reactions [45,46]. For this reason, more attention should be paid to implementing TXTL reactions in liposomes of defined size and dispersity.…”

“…encapsulated Xenopus egg extract in microdroplets of various size. The authors showed that the extract could undergo several mitotic oscillations, and described the influence of the size of the compartments on the period [45]. Sakamoto et al .…”