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Title: Effective RNA Regulation by Combination of Multiple Programmable RNA-Binding Proteins
Authors: Sugimoto, Misaki
Suda, Akiyo
Futaki, Shiroh
Imanishi, Miki  kyouindb  KAKEN_id  orcid (unconfirmed)
Author's alias: 須田, 明代
二木, 史朗
今西, 未来
Keywords: RNA binding protein
RNA regulation
Issue Date: 1-Oct-2020
Publisher: MDPI AG
Journal title: Applied Sciences
Volume: 10
Issue: 19
Thesis number: 6803
Abstract: RNAs play important roles in gene expression through translation and RNA splicing. Regulation of specific RNAs is useful to understand and manipulate specific transcripts. Pumilio and fem-3 mRNA-binding factor (PUF) proteins, programmable RNA-binding proteins, are promising tools for regulating specific RNAs by fusing them with various functional domains. The key question is: How can PUF-based molecular tools efficiently regulate RNA functions? Here, we show that the combination of multiple PUF proteins, compared to using a single PUF protein, targeting independent RNA sequences at the 3′ untranslated region (UTR) of a target transcript caused cooperative effects to regulate the function of the target RNA by luciferase reporter assays. It is worth noting that a higher efficacy was achieved with smaller amounts of each PUF expression vector introduced into the cells compared to using a single PUF protein. This strategy not only efficiently regulates target RNA functions but would also be effective in reducing off-target effects due to the low doses of each expression vector.
Rights: © 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 (
DOI(Published Version): 10.3390/app10196803
Appears in Collections:Journal Articles

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