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

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...

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Autores principales: Lin, Jyun-Liang, Ekas, Holly, Deaner, Matthew, Alper, Hal S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: KeAi Publishing 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6378893/
https://www.ncbi.nlm.nih.gov/pubmed/30820479
http://dx.doi.org/10.1016/j.synbio.2019.02.001
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author Lin, Jyun-Liang
Ekas, Holly
Deaner, Matthew
Alper, Hal S.
author_facet Lin, Jyun-Liang
Ekas, Holly
Deaner, Matthew
Alper, Hal S.
author_sort Lin, Jyun-Liang
collection PubMed
description 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.
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spelling pubmed-63788932019-02-28 CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering Lin, Jyun-Liang Ekas, Holly Deaner, Matthew Alper, Hal S. Synth Syst Biotechnol Article 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. KeAi Publishing 2019-02-15 /pmc/articles/PMC6378893/ /pubmed/30820479 http://dx.doi.org/10.1016/j.synbio.2019.02.001 Text en © 2019 Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Lin, Jyun-Liang
Ekas, Holly
Deaner, Matthew
Alper, Hal S.
CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
title CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
title_full CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
title_fullStr CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
title_full_unstemmed CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
title_short CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering
title_sort crispr-pin: modifying gene position in the nucleus via dcas9-mediated tethering
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6378893/
https://www.ncbi.nlm.nih.gov/pubmed/30820479
http://dx.doi.org/10.1016/j.synbio.2019.02.001
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