Reference - Detail
| RRC ID | 58787 |
|---|---|
| Author | Morita S, Horii T, Kimura M, Hatada I. |
| Title | Synergistic Upregulation of Target Genes by TET1 and VP64 in the dCas9-SunTag Platform. |
| Journal | Int J Mol Sci |
| Abstract |
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. |
| Volume | 21(5) |
| Published | 2020-2-25 |
| DOI | 10.3390/ijms21051574 |
| PII | ijms21051574 |
| PMID | 32106616 |
| PMC | PMC7084704 |
| MeSH | A549 Cells Basic-Leucine Zipper Transcription Factors / genetics Basic-Leucine Zipper Transcription Factors / metabolism CRISPR-Associated Protein 9 / genetics CRISPR-Associated Protein 9 / metabolism CRISPR-Cas Systems* Genetic Engineering / methods* Humans Mixed Function Oxygenases / genetics Mixed Function Oxygenases / metabolism Promoter Regions, Genetic Proto-Oncogene Proteins / genetics Proto-Oncogene Proteins / metabolism RNA, Guide, Kinetoplastida / genetics RNA, Guide, Kinetoplastida / metabolism Single-Chain Antibodies / genetics Single-Chain Antibodies / metabolism Up-Regulation* |
| IF | 4.556 |
| Times Cited | 0 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | A549(RCB0098) |