1. CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
- Author
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Jyun-Liang Lin, Holly Ekas, Matthew Deaner, and Hal S. Alper
- Subjects
Biotechnology ,TP248.13-248.65 ,Biology (General) ,QH301-705.5 - 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. Keywords: CRISPR, Chromosome organization, Chromosome biology, Gene positioning, Synthetic biology
- Published
- 2019
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