Combining live imaging, modelling and molecular genetics to understand transcriptional mechanism

为了生物体的正常发育和功能，基因必须在正确的地方和正确的时间被激活。在基因没有被正确开启，或者基因没有在正确的时间内开启的情况下，细胞功能丧失，这可能导致毁灭性的人类疾病，如癌症、神经变性和发育障碍。为了研究基因的开启和关闭是如何被控制的，我们使用了最先进的技术，直接观察活细胞中的单个基因。使用显微镜，我们可以观察细胞并评估特定基因是否开启或关闭。我们测量开、关周期的数量和持续时间，以及开相强度随时间的变化。我们测试当细胞受到刺激时，开关特性是如何变化的，以及它们是如何随着细胞的发育而随时间变化的。我们将这些观察结果与基因行为的简单计算机模拟进行比较，这些模拟是根据我们确定的规则开发的。当计算机无法模仿真实基因的开/关行为时，我们就知道我们的规则是错误的，需要改变。当我们有一套似乎起作用的规则时，我们就对实际的基因进行修改，然后观察并记录它们的行为是如何变化的。我们制定的规则也应该以一种可预测的方式适用于修改后的基因，所以如果它们不适用，我们就知道这些规则仍然是错误的，我们需要进一步评估它们。调整和完善这套规则的过程迫使我们不断发展对基因如何被控制的更全面的理解

For normal development and functioning of organisms, genes must be switched on in the correct place and at the correct time. In contexts where genes are not switched on correctly, or where genes are not on for the correct period of time, loss of cell function results, which can lead to devastating human diseases, such as cancer, neurodegeneration and developmental disorders. To study how the switching on and off of genes is controlled, we use a state-of-the-art technique, to directly watch single genes in living cells. Using a microscope, we can observe cells and assess whether a specific gene is on or off. We measure the number and duration of on and off periods and the variation in the strength of the on phase over time. We test how the on/off properties alter when cells are stimulated, and how they change over time as the cells develop.We compare these observations to simple computer simulations of gene behaviour, which are developed using rules we decide upon. When the computer fails to mimic the on/off behaviour of real genes, we learn that our rules are wrong, and need to be changed. When we have a set of rules which appears to work, we then make modifications to the actual genes, then watch and record how their behaviour changes. The rules we have developed should also apply in a predictable way to the modified gene, so if they do not, we know the rules are still wrong, and we need to further evaluate them. The process of adjusting and improving the set of rules forces us to continually develop a more complete understanding of how genes are controlled