Single-molecule investigations of fast biomolecular search processes initiating DNA repair

我的研究计划的总体目标是建立一个新的机制的快速分子搜索过程的理解，这是一个主机的关键细胞过程的基础。一类快速分子搜索过程，由于有限的分析技术，在很大程度上仍然没有表征，是在电离辐射、自由基和复制期间切割DNA后启动DNA修复的恒定过程的搜索。 现有的显微镜技术是不能够表征这些分子的搜索在生理条件下，因为它们不能同时解决：弱蛋白质-DNA和DNA-DNA相互作用;延长搜索轨迹的个别分子在几秒钟内，和毫秒的时间尺度的相互作用动力学;也不是他们与拓扑结构复杂或应变的DNA基板兼容。我将使用一套新的定制单显微镜技术来规避这些限制，这些技术扩展了CLIC成像。

The overarching objective of my research program is to create a new mechanistic understanding of the fast molecular search processes, which underlie a host of key cellular processes. One class of fast molecular search processes, that remain largely uncharacterized due to limited analysis techniques, is the search that initiates the constant processes of DNA repair following its cleavage by ionizing radiation, free radicals and during replication. Existing microscopy techniques are not capable of characterizing these molecular searches under physiological conditions because they can not simultaneously resolve: weak protein-DNA and DNA-DNA interactions; extended search trajectories of individual molecules over several seconds, and millisecond-timescale interaction kinetics; nor are they compatible with topologically complex or strained DNA substrates. I will circumvent these limitations using a new suite of custom single-microscopy techniques which extend CLIC-imaging.