Functional analysis and targeted manipulation of chromatin dynamics on mammalian rRNA genes and Nucleolar Organizer Regions (NORs) (A35*)

核糖体rna水平及其结果是蛋白质翻译速率在发育过程中发生变化，并且在不同细胞类型之间差异显著。然而，尽管它们很重要，但人们对rRNA基因如何受染色质机制调节知之甚少。我们最近开发了一种策略，允许使用gRNA多路复用和dCas9方法在体外和体内靶向操纵rDNA染色质。现在，我们将使用这种策略来操纵一些rDNA染色质组分（UBF， PARP， FACT）和机制（DNA甲基化，核小体重塑，非编码转录，代谢信号传导），以研究哪些是决定rRNA表达，细胞蛋白翻译率和哺乳动物干细胞行为的因果关系。

Levels of ribosomal RNAs and, as a consequence, rates of protein translation change during development and differ remarkably across cell types. Despite their importance however, very little is known about how rRNA genes are regulated by chromatin mechanisms. We have recently developed a strategy allowing targeted manipulation of rDNA chromatin in vitro and in vivo using gRNA multiplexing and dCas9 approaches. Now, we will use this strategy to manipulate a number of rDNA chromatin components (UBF, PARP, FACT) and mechanisms (DNA methylation, nucleosome remodelling, noncoding transcription, metabolic signalling), to investigate, which of those are causally determining rRNA expression, cellular protein translation rates and mammalian stem cell behaviour.