Administrative Supplement: Dynamics and evolution of synthetic and natural gene regulatory networks

项目摘要：NIGMS MIRA R35 GM 122561的行政补充 父项目名称：合成和天然基因调控网络的动力学和进化 本管理补充文件基于NOT-GM-21-030《特别关注通知（NOSI）： NIGMS R 01、R35和R37获奖者设备采购的行政补充”。是 瑞士生物技术公司Cytosurge AG请求部分支持FluidFM Bio系统。 FluidFM系统具有独特的重复、单个活细胞特异性、直接细胞内 细胞质或核内流体输送和提取，以及非破坏性单细胞 操纵这些能力将在促进母基金的进展方面发挥变革性作用， 其目的是了解基因网络动力学和随机性如何影响单细胞，从而影响细胞 人口。为了实现这一点，我们提出使用合成基因网络来生成特定的基因， 作为自然基因网络信号的空间和时间表达模式， 通过计算机建模对细胞群体行为和进化的后续影响， 实验进化FluidFM系统将加速合成基因的稳定基因组插入， 通过CRISPR、重组酶和遗传有效载荷的偶联核内递送， 单个细胞，通过细胞分离克隆生长，并通过从工程化的细胞中反复进行“活检”， 研究单个活细胞的转录组和蛋白质组网络如何对扰动做出反应 通过合成基因电路。总体而言，FluidFM系统将提供独特的能力， 复杂的网络如何在生物学中实现跨时空尺度的控制，从分子到 细胞解决这些问题将教会我们如何控制适应性细胞群， 与理解、预测和可能预防癌症和微生物耐药性有关。

Project Summary: Administrative Supplement to NIGMS MIRA R35GM122561 Parent project title: Dynamics and evolution of synthetic and natural gene regulatory networks This Administrative Supplement is based on NOT-GM-21-030, “Notice of Special Interest (NOSI): Administrative Supplements for Equipment Purchases for NIGMS R01, R35, and R37 Awardees”. It is a request for partial support of a FluidFM Bio system by Cytosurge AG, a Swiss Biotech company. The FluidFM system has unique capabilities for repeated, single live cell-specific, direct intra- cytoplasmic or intra-nuclear delivery and extraction of fluids, as well as nondestructive single cell manipulation. These capabilities will be transformative in promoting progress with the parent grant, which aims to learn how gene network dynamics and stochasticity affect single cells and thereby cell populations. To achieve this, we proposed using synthetic gene networks to generate specific gene expression patterns in space and time that serve as signals for natural gene networks, studying the subsequent effects on cell population behavior and evolution by computational modeling and experimental evolution. The FluidFM system will accelerate the stable genomic insertion of synthetic gene circuits by coupled intranuclear delivery of CRISPR, recombinase and genetic payloads into single cells, by cell isolation for clonal outgrowth, and by repeated “biopsies” taken from engineered single living cells to study how their transcriptomic and proteomic networks respond to perturbations by synthetic gene circuits. Overall, the FluidFM system will provide unique capabilities to illuminate how complex networks enable control across scales of space and time in biology, from molecules to cells. Addressing these questions will teach us how to control adapting cell populations, which is relevant for understanding, predicting and possibly preventing cancer and microbial drug resistance.