Cellular Phenotype sequencing (CePh-seq): Decoding the morphological and molecular age-related phenotypes of single cells

我们体内的所有细胞都有相同的遗传信息，但拥有相同基因组的不同细胞表现出非常不同的特性，做着非常不同的事情。这些差异在很大程度上归结为每个细胞中哪些基因实际上是被激活的。人们还发现，具有相同转录基因组合的细胞通常看起来非常相似，行为方式也很相似，但理解活跃基因和外观之间的联系是非常困难的，因为没有自动方法来收集看起来都相似的细胞。在这个BBSRC项目中，我们将开发一种新的生物技术仪器，它可以采集混合的细胞样本，准确地测量它们的形态特征，然后选择那些具有特定形态的细胞，用唯一的分子识别符对它们进行条形码编码，最后测量它们所有基因的活性。条形码的使用是必不可少的，因为分析基因的过程破坏了细胞，只观察细胞内的分子。通过将这种分子条形码与细胞被摧毁前的图像联系起来，将有可能调查特定基因和细胞外观之间的关系。我们将使用这项新技术来研究衰老中迄今尚未解决的问题，这可能会揭示老细胞如何包装他们的DNA，以及这如何改变活跃的基因。特别是，我们将研究细胞衰老时细胞核的变化--细胞停止分裂的过程与衰老和癌症有关。已经观察到，一些衰老细胞中的DNA可以重排形成独特的亮点模式，但这种重排的原因和功能仍不清楚，很难研究，因为只有一些细胞表现出这种变化。通过根据细胞DNA的排列分离细胞，我们将能够研究这些细胞与其同伴的不同之处，这可能为理解衰老过程中细胞的变化提供关键的见解。

All the cells in our body have the same genetic information, yet different cells which have the same genome display very different properties and do very different things. A large part of these differences comes down to which genes are actually turned on within each cell. It has also been seen that cells which have the same combination of genes being transcribed often look very similar and behave in a similar way, yet understanding the connection between active genes and appearance is very difficult, as there is no automated way to collect cells which all look similar. In this BBSRC project, we will develop a new biotechnology instrument which can take a mixed sample of cells, accurately measure their morphological properties, then select those cells with a particular morphology of interest, barcode them with a unique molecular identifier, and finally measure the activity of all of their genes. The use of the barcode is essential because the process of analysing the genes destroys the cells and only looks at the molecules within the cell. By linking this molecular barcode to the image of the cell before it was destroyed, it will be possible to investigate the relationship between particular genes and how cells look.We will use this new technology to investigate a so far unsolvable problem in ageing which may shed light on how old cells pack their DNA and how this can change the active genes. In particular, we will investigate the changes in the nuclei of cells when they become senescent - a process by which cells stop dividing which is linked to ageing and cancer. It has been observed that the DNA in some senescent cells can rearrange to form a distinctive pattern of bright spots, however the causes and function of such rearrangements are still unknown and difficult to investigate as it is only some of the cells which display the change. By isolating cells based on the arrangement of their DNA, we will be able to investigate what makes these cells different from their companions, which may provide critical insight into understanding the cellular changes of growing old.