CRISPR-Cas-Mediated Tethering Recruits the Yeast HMR Mating-Type Locus to the Nuclear Periphery but Fails to Silence Gene Expression

To investigate the relationship between genome structure and function, we have developed a programmable CRISPR-Cas system for nuclear peripheral recruitment in yeast. We benchmarked this system at the HMR and GAL2 loci, both of which are well-characterized model systems for localization to the nuclear periphery. Using microscopy and gene silencing assays, we demonstrate that CRISPR-Cas-mediated tethering can recruit the HMR locus but does not detectably silence reporter gene expression. A previously reported Gal4-mediated tethering system does silence gene expression, and we demonstrate that the silencing effect has an unexpected dependence on the properties of the protein tether. The CRISPR-Cas system was unable to recruit GAL2 to the nuclear periphery. Our results reveal potential challenges for synthetic genome structure perturbations and suggest that distinct functional effects can arise from subtle structural differences in how genes are recruited to the periphery.

Automated summary

Research on the relationship between genome structure and function using a programmable CRISPR-Cas system in yeast showed that tethering the HMR locus did not significantly silence gene expression. A comparison with the Gal4 system revealed that the silencing effect is unexpectedly dependent on the properties of the protein tether. Additionally, the CRISPR-Cas system was unable to recruit the GAL2 locus to the nuclear periphery, suggesting potential challenges in synthetic genome structure perturbations.