CTCF control of genome stability in ageing

Objetivo

. Genome stability is one of the most important features in maintaining tissue homeostasis throughout the human lifespan. The research presented here will dissect how the insulator protein CCCTC-binding factor (CTCF), a ubiquitous 11 zinc finger transcription factor, controls the stability of the mammalian genome during ageing.
. In Aim 1, we will elucidate how CTCF and tissue-specific master regulators maintain the functional stability of the genome during healthy ageing by developing a novel protocol to map simultaneously transcription and open chromatin in isolated hepatocyte nuclei. Using this protocol, we will explore how CTCF binding stabilizes cellular homeostasis during ageing by knocking down CTCF in vivo, both in isolation and simultaneously with knock down of liver-specific master regulators.
. In Aim 2, we will reveal the molecular mechanisms underlying CTCF binding sites as susceptibility loci for somatic mutations. We will profile the mutations in open chromatin of single nuclei immediately following acute exposure to a chemical mutagen; comparing how the pattern of mutations in CTCF bound regions changes across an allelic series of CTCF knockdown mice will reveal how CTCF binding shapes the stability of the genome towards mutations.
. These integrated strategies develop and deploy powerful, cutting-edge experimental approaches to reveal novel aspects of how CTCF binding stabilises the mammalian genome during healthy ageing as well as during mutagenesis.