Cellular logic of phenotype

表型的细胞逻辑 这个应用程序的目标是开发一种可预测和 在培养中可重复性地改变原代细胞的表型。分化的细胞类型 它们的RNA图谱不同(相对丰度和绝对丰度 它们表达的RNA)。我假设，通过将整个RNA图谱从 以一种使受体细胞存活依赖于 供体RNA，供体RNA会将受体改变为目的表型， 模仿供体细胞的表型。这一过程被称为转录组诱导 表型重构(TIPeR)。有能力将细胞表型在 细胞将为控制细胞的机制提供重要的新见解 差异化。实现这一目标的理论和技术策略正在 在我的实验室里研发出来的。具体地说，利用激光诱导光转基因 (在我的实验室开发)，我们在宿主初级细胞中瞬时产生毛孔，通过 哪些RNA种群(其中RNA的种类和丰度受到严格控制)， 可以扩散。初步数据显示，供体细胞的RNA群体携带着“记忆” 功能“，即供体RNA可以诱导基因组转录的长期变化 宿主细胞从而将宿主细胞的功能表型改变为 目的表型。这在一定程度上是由于特定的 由宿主细胞RNA混合物制成的蛋白质。通过发展各种高技术， 与TIPeR相结合的生产能力定量“OMICS”级表型技术 预计表型的“基因组学逻辑”将被识别出来。一个 对这一逻辑的理解将允许随意创建特定的细胞类型。一种能力 有选择地和合理地创造细胞表型有望提供重要的 对细胞极性、功能和基本机制的洞察 表型稳定，并可能产生新的“个体化药物疗法”。

The Cellular Logic of Phenotype The goal of this application is to develop a strategy for predictably and reproducibly altering the phenotype of primary cells in culture. Differentiated cell types differ from each other in their RNA profiles (relative as well as absolute abundances of the RNAs they express). I hypothesize that, by the transferring entire RNA profiles from donor to recipient cells in a way that makes the recipient cells' survival dependent on donor RNA, the donor RNA will change the recipient into a destination phenotype that mimics the donor cell phenotype. This procedure is called Transcriptome Induced Phenotype Remodeling (TIPeR). Having the ability to transfer cell phenotypes between cells would provide important new insights into mechanisms controlling cell differentiation. The theory and technical strategies to accomplish this are being developed in my laboratory. Specifically, using laser light induced phototransfection (developed in my lab), we transiently produce pores in the host primary cell, through which RNA populations (in which RNA species and abundances are carefully controlled), can diffuse. Preliminary data shows that donor cell RNA populations carry "memory functions" in that, donor RNA can induce long-term changes in genomic transcription of the host cells thereby changing the functional phenotype of the host cells to that of the destination phenotype. This is due in part to the activity and abundances of the specific proteins made from the host cell RNA mixture. Through developing various high- throughput quantitative "Omics" level phenotyping technologies coupled with the TIPeR procedure it is anticipated that the "genomics logic" of phenotype will be discerned. An understanding of this logic will permit the creation of specific cell types at will. The ability to selectively and rationally create cellular phenotypes promises to provide important insights into the fundamental mechanisms underlying cellular polarity, functioning and phenotype stability and may yield novel "individualized medicinal therapeutics".