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Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform
Overexpression of a gene of interest is a general approach used in both basic research and therapeutic applications. However, the conventional approach involving overexpression of exogenous genes has difficulty achieving complete genome coverage, and is also limited by the cloning capacity of viral...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7084704/ https://www.ncbi.nlm.nih.gov/pubmed/32106616 http://dx.doi.org/10.3390/ijms21051574 |
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author | Morita, Sumiyo Horii, Takuro Kimura, Mika Hatada, Izuho |
author_facet | Morita, Sumiyo Horii, Takuro Kimura, Mika Hatada, Izuho |
author_sort | Morita, Sumiyo |
collection | PubMed |
description | Overexpression of a gene of interest is a general approach used in both basic research and therapeutic applications. However, the conventional approach involving overexpression of exogenous genes has difficulty achieving complete genome coverage, and is also limited by the cloning capacity of viral vectors. Therefore, an alternative approach would be to drive the expression of an endogenous gene using an artificial transcriptional activator. Fusion proteins of dCas9 and a transcription activation domain, such as dCas9–VP64, are widely used for activation of endogenous genes. However, when using a single sgRNA, the activation range is low. Consequently, tiling of several sgRNAs is required for robust transcriptional activation. Here we describe the screening of factors that exhibit the best synergistic activation of gene expression with TET1 in the dCas9–SunTag format. All seven factors examined showed some synergy with TET1. Among them, VP64 gave the best results. Thus, simultaneous tethering of VP64 and TET1 to a target gene using an optimized dCas9–SunTag format synergistically activates gene expression using a single sgRNA. |
format | Online Article Text |
id | pubmed-7084704 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-70847042020-03-24 Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform Morita, Sumiyo Horii, Takuro Kimura, Mika Hatada, Izuho Int J Mol Sci Communication Overexpression of a gene of interest is a general approach used in both basic research and therapeutic applications. However, the conventional approach involving overexpression of exogenous genes has difficulty achieving complete genome coverage, and is also limited by the cloning capacity of viral vectors. Therefore, an alternative approach would be to drive the expression of an endogenous gene using an artificial transcriptional activator. Fusion proteins of dCas9 and a transcription activation domain, such as dCas9–VP64, are widely used for activation of endogenous genes. However, when using a single sgRNA, the activation range is low. Consequently, tiling of several sgRNAs is required for robust transcriptional activation. Here we describe the screening of factors that exhibit the best synergistic activation of gene expression with TET1 in the dCas9–SunTag format. All seven factors examined showed some synergy with TET1. Among them, VP64 gave the best results. Thus, simultaneous tethering of VP64 and TET1 to a target gene using an optimized dCas9–SunTag format synergistically activates gene expression using a single sgRNA. MDPI 2020-02-25 /pmc/articles/PMC7084704/ /pubmed/32106616 http://dx.doi.org/10.3390/ijms21051574 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Communication Morita, Sumiyo Horii, Takuro Kimura, Mika Hatada, Izuho Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform |
title | Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform |
title_full | Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform |
title_fullStr | Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform |
title_full_unstemmed | Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform |
title_short | Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9–SunTag Platform |
title_sort | synergistic upregulation of target genes by tet1 and vp64 in the dcas9–suntag platform |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7084704/ https://www.ncbi.nlm.nih.gov/pubmed/32106616 http://dx.doi.org/10.3390/ijms21051574 |
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