CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering

Synthetic and Systems Biotechnology
Jyun-Liang LinHal S Alper

Abstract

Spatial organization of DNA within the nucleus is important for controlling DNA replication and repair, genetic recombination, and gene expression. Here, we present CRISPR-PIN, a CRISPR/dCas9-based tool that allows control of gene Position in the Nucleus for the yeast Saccharomyces cerevisiae. This approach utilizes a cohesin-dockerin interaction between dCas9 and a perinuclear protein. In doing so, we demonstrate that a single gRNA can enable programmable interaction of nuclear DNA with the nuclear periphery. We demonstrate the utility of this approach for two applications: the controlled segregation of an acentric plasmid and the re-localization of five endogenous loci. In both cases, we obtain results on par with prior reports using traditional, more cumbersome genetic systems. Thus, CRISPR-PIN offers the opportunity for future studies of chromosome biology and gene localization.

Citations

Feb 10, 2021·Nature Reviews. Genetics·Haifeng WangLei S Qi

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Methods Mentioned

BETA
PCR
confocal microscopy

Software Mentioned

ImageJ
SAS
Zen
IDT

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