General applicability of synthetic gene-overexpression for cell-type ratio control via reprogramming
General applicability of synthetic gene-overexpression for cell-type ratio control via reprogramming
复制标题
合成基因过表达通过重编程控制细胞类型比例的一般适用性
DOI：
10.1021/sb400102w
复制
发表时间：
2013
影响因子：
4.7
通讯作者：
K. Ishimatsu
中科院分区：
文献类型：
--
作者：
吉田 由紀子;陶山 史朗;山本 裕紹;Ken Komiya and Masayuki Yamamura;K. Ishimatsu
多稳态系统中细胞类型比例的控制需要对细胞初始状态进行大范围的控制。这里，使用 E 中的合成电路。在大肠杆菌中，我们描述了使用简单的基因过表达系统与双稳态切换开关相结合，以实现细胞状态的大范围控制，从而生成任意细胞类型比率。理论上，过表达诱导暂时将双稳态系统改变为单稳态系统，其中可以通过调节过表达水平在大范围内操纵细胞的单一稳态的位置。这种诱导的细胞状态在过度表达停止后成为基础双稳态系统的初始状态，从而恢复原始的双稳态系统。我们通过实验证明，过度表达诱导单峰细胞分布，随后过度表达的撤销产生双峰分布。此外，按照理论上的设计，通过调节小分子的浓度来调节过度表达水平，可以产生任意的细胞类型比例。
Control of the cell-type ratio in multistable systems requires wide-range control of the initial states of cells. Here, using a synthetic circuit inE. coli, we describe the use of a simple gene-overexpression system combined with a bistable toggle switch, for the purposes of enabling the wide-range control of cellular states and thus generating arbitrary cell-type ratios. Theoretically, overexpression induction temporarily alters the bistable system to a monostable system, in which the location of the single steady state of cells can be manipulated over a wide range by regulating the overexpression levels. This induced cellular state becomes the initial state of the basal bistable system upon overexpression cessation, which restores the original bistable system. We experimentally demonstrated that the overexpression induced a monomodal cell distribution, and subsequent overexpression withdrawal generated a bimodal distribution. Furthermore, as designed theoretically, regulating the overexpression levels by adjusting the concentrations of small molecules generated arbitrary cell-type ratios.