Synthetic memory circuits for tracking human cell fate
Synthetic memory circuits for tracking human cell fate
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DOI：
10.1101/gad.189035.112
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发表时间：
2012-07-01
影响因子：
10.5
通讯作者：
Silver, Pamela A.
中科院分区：
文献类型：
--
作者：
Burrill, Devin R.;Inniss, Mara C.;Silver, Pamela A.
从疾病进展到干细胞分化的各种生物现象的典型特征是细胞对短暂的环境提示的长期反应。虽然在生物学上相关，但这些长期反应的异质性很难在种群水平上进行评估，因此有必要开发生物学工具来跟踪亚种群内的细胞命运。在这里，我们提出了一种新的合成生物学方法来识别和跟踪哺乳动物细胞亚群。我们构建了三个基因集成电路，它们使用双稳态自动调节转录反馈来保留对短暂刺激的记忆。这些“记忆装置”被用来分离和跟踪对多西环素、低氧或DNA损伤剂有不同反应的细胞的后代。在低氧或紫外线照射后，反应强烈的细胞激活记忆装置，并在最初刺激后的多代中表现出基因表达、生长速度和生存能力的变化。综上所述，这些结果表明，在长期细胞行为不同的亚群中存在对低氧和DNA损伤的可遗传记忆。
A variety of biological phenomena, from disease progression to stem cell differentiation, are typified by a prolonged cellular response to a transient environmental cue. While biologically relevant, heterogeneity in these long-term responses is difficult to assess at the population level, necessitating the development of biological tools to track cell fate within subpopulations. Here we present a novel synthetic biology approach for identifying and tracking mammalian cell subpopulations. We constructed three genomically integrated circuits that use bistable autoregulatory transcriptional feedback to retain memory of exposure to brief stimuli. These "memory devices'' are used to isolate and track the progeny of cells that responded differentially to doxycycline, hypoxia, or DNA-damaging agents. Following hypoxic or ultraviolet radiation exposure, strongly responding cells activate the memory device and exhibit changes in gene expression, growth rates, and viability for multiple generations after the initial stimulus. Taken together, these results indicate that a heritable memory of hypoxia and DNA damage exists in subpopulations that differ in long-term cell behavior.