Programming the Notch response: optogenetic strategies to manipulate pathway activity

布景主题：Notch信号通路是少数协调发育的细胞信号通路之一，调节在许多发育环境中形成的细胞的类型和数量。它的功能包括维持干细胞/祖细胞、调节细胞命运、组织生长模式等等。此外，异常的Notch活性与包括癌症在内的疾病有关。实验室工作的一个主要重点是了解Notch通路是如何运作的，以及是什么使其产生不同的功能结果。最近，我们开发了一些方法，使我们能够在活细胞中跟踪特定的靶基因，从而有可能研究核事件，包括转录因子动力学和染色质环境如何影响基因靶点的选择。剩下的挑战是如何高精度地激活Notch。为了实现这一点，我们将采用光遗传学方法，通过将其融合到光敏结构域来释放Notch受体的活性形式。最终，这些光遗传学工具与跟踪途径中其他组分行为的方法相结合，将用于生成精确的模型，以了解活性Notch如何在细胞内运作以靶向正确的基因，以及如何通过致病修饰破坏这些基因。

Theme: Understanding the rules of lifeThe Notch pathway is one of a small handful of cell signalling pathways that coordinate development, regulating the types and numbers of cells formed in many developmental contexts. Its functions include maintenance of stem/progenitor cells, regulation of cell fates, organizing patterns of growth, and many others. In addition, aberrant Notch activity is implicated in diseases including cancers. A major focus of the work in the lab is on understanding how the Notch pathway operates and what enables its different functional outcomes.Recently we have developed methods that allow us to follow specific target genes in living cells making it possible to investigate nuclear events including how the transcription factor dynamics and the chromatin context influences the selection of gene targets. A remaining challenge is how to activate Notch with high precision. To achieve this we will adopt optogenetic methods to release an active form of the Notch receptor, by fusing it to light sensitive domains. Ultimately, these optogenetic tools, combined with methods to track the behaviour of other components in the pathway, will be used to generate accurate models of how the active Notch operates inside a cell to target the correct genes and how this can be disrupted by disease causing modifications.