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The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice

We report here a robust, tunable, and reversible transcription control system for endogenous genes. The REMOTE-control system (Reversible Manipulation of Transcription at Endogenous loci) employs enhanced lac repression and tet activation systems. With this approach, we show in mouse embryonic stem...

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Autores principales: Lee, Kwang-Ho, Oghamian, Shirley, Park, Jin-A, Kang, Liang, Laird, Peter W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716148/
https://www.ncbi.nlm.nih.gov/pubmed/28981717
http://dx.doi.org/10.1093/nar/gkx829
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author Lee, Kwang-Ho
Oghamian, Shirley
Park, Jin-A
Kang, Liang
Laird, Peter W.
author_facet Lee, Kwang-Ho
Oghamian, Shirley
Park, Jin-A
Kang, Liang
Laird, Peter W.
author_sort Lee, Kwang-Ho
collection PubMed
description We report here a robust, tunable, and reversible transcription control system for endogenous genes. The REMOTE-control system (Reversible Manipulation of Transcription at Endogenous loci) employs enhanced lac repression and tet activation systems. With this approach, we show in mouse embryonic stem cells that endogenous Dnmt1 gene transcription could be up- or downregulated in a tunable, inducible, and reversible manner across nearly two orders of magnitude. Transcriptional repression of Dnmt1 by REMOTE-control was potent enough to cause embryonic lethality in mice, reminiscent of a genetic knockout of Dnmt1 and could substantially suppress intestinal polyp formation when applied to an Apc(Min) model. Binding by the enhanced lac repressor was sufficiently tight to allow strong attenuation of transcriptional elongation, even at operators located many kilobases downstream of the transcription start site and to produce invariably tight repression of all of the strong viral/mammalian promoters tested. Our approach of targeting tet transcriptional activators to the endogenous Dnmt1 promoter resulted in robust upregulation of this highly expressed housekeeping gene. Our system provides exquisite control of the level, timing, and cell-type specificity of endogenous gene expression, and the potency and versatility of the system will enable high resolution in vivo functional analyses.
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spelling pubmed-57161482017-12-08 The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice Lee, Kwang-Ho Oghamian, Shirley Park, Jin-A Kang, Liang Laird, Peter W. Nucleic Acids Res Gene regulation, Chromatin and Epigenetics We report here a robust, tunable, and reversible transcription control system for endogenous genes. The REMOTE-control system (Reversible Manipulation of Transcription at Endogenous loci) employs enhanced lac repression and tet activation systems. With this approach, we show in mouse embryonic stem cells that endogenous Dnmt1 gene transcription could be up- or downregulated in a tunable, inducible, and reversible manner across nearly two orders of magnitude. Transcriptional repression of Dnmt1 by REMOTE-control was potent enough to cause embryonic lethality in mice, reminiscent of a genetic knockout of Dnmt1 and could substantially suppress intestinal polyp formation when applied to an Apc(Min) model. Binding by the enhanced lac repressor was sufficiently tight to allow strong attenuation of transcriptional elongation, even at operators located many kilobases downstream of the transcription start site and to produce invariably tight repression of all of the strong viral/mammalian promoters tested. Our approach of targeting tet transcriptional activators to the endogenous Dnmt1 promoter resulted in robust upregulation of this highly expressed housekeeping gene. Our system provides exquisite control of the level, timing, and cell-type specificity of endogenous gene expression, and the potency and versatility of the system will enable high resolution in vivo functional analyses. Oxford University Press 2017-12-01 2017-09-14 /pmc/articles/PMC5716148/ /pubmed/28981717 http://dx.doi.org/10.1093/nar/gkx829 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Gene regulation, Chromatin and Epigenetics
Lee, Kwang-Ho
Oghamian, Shirley
Park, Jin-A
Kang, Liang
Laird, Peter W.
The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice
title The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice
title_full The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice
title_fullStr The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice
title_full_unstemmed The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice
title_short The REMOTE-control system: a system for reversible and tunable control of endogenous gene expression in mice
title_sort remote-control system: a system for reversible and tunable control of endogenous gene expression in mice
topic Gene regulation, Chromatin and Epigenetics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5716148/
https://www.ncbi.nlm.nih.gov/pubmed/28981717
http://dx.doi.org/10.1093/nar/gkx829
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