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

项目总结：NIGMS Mira R35GM122561的行政补充 父项目标题：合成和天然基因调控网络的动态和进化 本行政补编基于NOT-GM-21-030《特别利益通知(NOSI)： NIGMS R01、R35和R37获奖者的设备采购行政补充“。它是 瑞士生物科技公司细胞清理股份公司对FluidFM Bio系统的部分支持请求。 FluidFM系统具有重复的、单个活细胞特定的、直接内照射的独特能力 细胞质或核内液体的输送和提取，以及非破坏性单细胞 操纵。这些能力将在促进父母赠款的进展方面产生变革性的作用， 旨在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产生特定的基因 作为自然基因网络信号的空间和时间表达模式，研究 通过计算模型和方法对细胞群体行为和进化的后续影响 实验进化论。FluidFM系统将加速合成的稳定基因组插入 CRISPR、重组酶和基因有效载荷耦合核内递送的基因电路 单个细胞，通过克隆生长的细胞分离，通过重复从工程细胞中提取的“活组织检查” 单个活细胞来研究它们的转录和蛋白质组网络如何对扰动做出反应 通过人工合成的基因电路。总体而言，FluidFM系统将提供独特的能力来照明 复杂的网络如何在生物学中实现跨越空间和时间尺度的控制，从分子到 细胞。解决这些问题将教会我们如何控制适应的细胞群体，这是 与了解、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.