Genetically encoded live cell sensors of chromatin state

项目摘要 在过去的半个世纪里，生物学家已经产生了大量的知识， 关于染色质（蛋白质和RNA的集合）如何调节基因的假说 表情这些知识有可能改变我们对发育和疾病的理解， 刺激新的治疗策略。为了实现这一潜力，需要新的实验工具来直接测试 现有的假说，并揭示染色质组分的功能。特别有用的是 直接和特异性地跟踪活细胞中染色质的生物化学和生物物理状态。这将 解锁我们研究染色质状态响应细胞扰动的动态变化的能力。观察 时间序列中的细胞内事件也将使功能研究能够证明基因的变化， 直接响应染色质状态变化的调节。为了实现这一目标，我们将开发小蛋白质 结合特定的生化染色质修饰。这些粘合剂将被基因编码， 它们在活细胞显微镜中的应用。这些染色质结合剂的发展将支持一个新的前沿， 染色质研究他们将建立在从染色质状态的静态图中获得的信息的基础上， 将我们对染色质的理解扩展到时间和空间两个维度。

Project Summary Over the past half-century, biologists have generated a critical mass of knowledge and many compelling hypotheses about how chromatin, a collection of proteins and RNA scaffolded to DNA, regulates gene expression. This knowledge has the potential to transform our understanding of development and disease and spur new therapeutic strategies. To realize this potential, new experimental tools are needed to directly test existing hypotheses and reveal the functions of chromatin components. Of particular use would be approaches that directly and specifically track the biochemical and biophysical states of chromatin in live cells. This would unlock our ability to study dynamic changes in chromatin state in response to cellular perturbations. Observing intracellular events in temporal sequence would also enable functional studies demonstrating changes in gene regulation in direct response to changes in chromatin state. Towards this goal, we will develop small proteins that bind specific biochemical chromatin modifications. These binders will be genetically encoded, enabling their use in live cell microscopy. The development of these chromatin binders will support a new frontier in chromatin research. They will build upon the information garnered from static maps of chromatin state and expand our understanding of chromatin into both the temporal and spatial dimensions.