Time and context-controlled depletion of proteins in single cells

我在这个补助金申请中提出了一个新的策略，以快速消耗蛋白质或蛋白质内的单域在一个精确定义的时间，无论是上下文相关或独立的方式。该策略将与两个过程的单细胞分析紧密相连：酵母中的早期细胞极化和胞质分裂。它将被用来最小化干扰蛋白质网络参与这两个过程中的任何一个，并定量测量这些扰动在单细胞中的后果。我们的目标是连接系统属性的蛋白质网络与执行的形态发生程序，更一般地说，以减少相互作用的数据（蛋白质或基因）的积累和理解这些相互作用的网络及其相关的生物过程的意义之间仍然不断增长的差距。因此，这一建议解决了一个重要的问题，在最近的一项调查中被确定为系统生物学取得实质性进展的一个要求。

I propose in this grant application a new strategy to rapidly deplete proteins or single domains within proteins at a precisely defined time in either a context-dependent or -independent manner. The strategy will be tightly linked to the single cell analysis of two processes: Early cell polarization and cytokinesis in yeast. It will be used to minimally perturb the network of proteins involved in either of the two processes and to quantitatively measure the consequences of these perturbations in single cells. The aim is to connect system properties of a protein network with the execution of a morphogenetic program, and, more generally, to reduce the still growing gap between the accumulation of interaction data (protein or gene) and the understanding of the significance of these interactions for the networks and their associated biological processes. This proposal thus addresses an important issue that was identified in a recent survey as one requirement for a substantial progress in systems biology1.