Dynamic Cell Programming with Quorum Sensing-Controlled CRISPRi Circuit.,ACS Synthetic Biology

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Dynamic Cell Programming with Quorum Sensing-Controlled CRISPRi Circuit.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-06-02 , DOI: 10.1021/acssynbio.0c00148
Yilan Liu ₁ , Jinjin Chen ₁ , David Crisante ₁ , Jhoselyn Marisol Jaramillo Lopez ₁ , Radhakrishnan Mahadevan _{1,

2}

Affiliation

Synthetic biology is enabling rapid advances in the areas of biomanufacturing and live therapeutics. Dynamic circuits that can be used to regulate cellular resources and microbial community behavior represent a defining focus of synthetic biology, and have attracted tremendous interest. However, the existing dynamic circuits are mostly gene editing-dependent or cell lysis-based, which limits their broad and convenient application, and in some cases, such lysis-based circuits can suffer from genetic instability due to evolution. There is limited research in quorum sensing-assisted CRISPRi, which can function in a gene editing-independent manner. Here, we constructed a series of quorum sensing controlled CRISPRi systems (Q-CRISPRi), which can dynamically program bacteria by using customized sgRNA without introducing cell lysis. We successfully applied Q-CRISPRi circuits to dynamically program gene expression, population density, phenotype, physical property, and community composition of microbial consortia. The strategies reported here represent methods for dynamic cell programming and could be effective in programming industrially and medically important microorganisms to offer better control of their metabolism and behavior.

中文翻译：

使用群体感应控制的 CRISPRi 电路进行动态细胞编程。

合成生物学正在推动生物制造和活体治疗领域的快速发展。可用于调节细胞资源和微生物群落行为的动态电路代表了合成生物学的一个明确焦点，并引起了极大的兴趣。然而，现有的动态电路大多依赖于基因编辑或基于细胞裂解，这限制了它们的广泛和方便的应用，并且在某些情况下，这种基于裂解的电路可能会因进化而遭受遗传不稳定。群体感应辅助 CRISPRi 可以以独立于基因编辑的方式发挥作用，目前的研究有限。在这里，我们构建了一系列群体感应控制的 CRISPRi 系统（Q-CRISPRi），该系统可以使用定制的 sgRNA 动态编程细菌，而无需引入细胞裂解。我们成功应用 Q-CRISPRi 电路对微生物群落的基因表达、种群密度、表型、物理特性和群落组成进行动态编程。这里报道的策略代表了动态细胞编程的方法，并且可以有效地对工业和医学上重要的微生物进行编程，以更好地控制它们的新陈代谢和行为。

更新日期：2020-06-19

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