Yeast cells cycle asynchronously, meaning that each cell in the population buds at a different time (Fig. Activation of the cyclin-dependent kinase (CDK) Cdc28 by any of the cyclins Cln1, Cln2 and Cln3 is necessary to overcome the START checkpoint 4, 5, 6 and to irreversibly enter the cell cycle. In yeast, this START checkpoint is found in the late G 1 phase. A red nuclear fluorescence reporter (Htb2-mCherry) and a yellow fluorescence reporter, consisting of a fusion protein between endogenous mitotic septin Cdc10 and YFP (Cdc10-YFP), are also present in this strain.ĭuring the G 1 phase, the cell can either commit to the cell cycle and enter the S phase, or if conditions are not favourable, arrest the cell cycle. An exogeneous copy of the G 1 cyclin gene CLN2 is placed under the control of the methionine-repressible promoter P MET3. Cells are deleted only for the G 1 cyclin Cln3, hence cells continuously cycle. A red fluorescent nuclear reporter, consisting of a fusion protein between the endogenous histone H2B protein and the mCherry (Htb2-mCherry) is also present in this strain. A yellow fluorescent protein (YFP) is expressed under the control of the endogenous promoter P CLN2. Cells can cycle only in the absence of methionine. Cells are deleted for genes encoding for the G 1 cyclins Cln1-3 while an exogeneous G 1 cyclin gene CLN2 is placed under the control of the methionine-repressible promoter P MET3. Schematics of the computer-controlled microfluidics platform to automatically synchronise the cell cycle across a population of yeast cells. b Yeast cells do not cycle synchronously in a population. Yeast strains were engineered to initiate the cell cycle upon methionine depletion from the growth medium (input). In addition to providing an avenue for yeast cell cycle synchronisation, our work shows that control engineering can be used to automatically steer complex biological processes towards desired behaviours similarly to what is currently done with robots and autonomous vehicles.Ī Schematic representation of the cell cycle in yeast Saccharomyces cerevisiae. Our work builds upon solid theoretical foundations provided by Control Engineering. The computer implements a controller algorithm to decide when, and for how long, to change the growth medium to synchronise the cell-cycle across the population.
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Here, we build a cyber-genetic system to achieve long-term synchronisation of the cell population, by interfacing genetically modified yeast cells with a computer by means of microfluidics to dynamically change medium, and a microscope to estimate cell cycle phases of individual cells. Although there are several experimental approaches to synchronise cells, these usually work only in the short-term. Yeast cells cycle asynchronously with each cell in the population budding at a different time.
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The cell cycle is the process by which eukaryotic cells replicate.