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

我们身体里所有的细胞都有相同的遗传信息，然而拥有相同基因组的不同细胞却表现出非常不同的特性，做着非常不同的事情。这些差异的很大一部分归结于每个细胞中哪些基因实际上是被激活的。我们还发现，具有相同转录基因组合的细胞通常看起来非常相似，行为方式也相似，但理解活性基因和外观之间的联系是非常困难的，因为没有自动收集看起来都很相似的细胞的方法。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性，然后选择那些具有特定形态的细胞，用独特的分子标识符对它们进行条形码，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会破坏细胞，只会看到细胞内的分子。通过将这种分子条形码与细胞被破坏前的图像联系起来，研究特定基因与细胞外观之间的关系将成为可能。我们将利用这项新技术来研究一个迄今为止无法解决的衰老问题，这可能会揭示老细胞是如何包装DNA的，以及这是如何改变活性基因的。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，这与衰老和癌症有关。据观察，一些衰老细胞的DNA可以重新排列，形成一个独特的亮点模式，然而，这种重新排列的原因和功能仍然是未知的，而且很难研究，因为只有一些细胞会显示这种变化。通过根据DNA的排列分离细胞，我们将能够研究是什么使这些细胞不同于它们的同伴，这可能为理解细胞变老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.