ID 110894
Author
Kojima, Takamitsu Tokushima University
Kinjo, Nozomi Tokushima University
Hashimoto, Yosuke Tokushima University
Furukawa, Kazuhiro Tokushima University
Keywords
4′-thioDNA
innate immune response
RNA interference
shRNA
TLR9
Content Type
Journal Article
Description
The development of a versatile technique to induce RNA interference (RNAi) without immune stimulation in vivo is of interest as existing approaches to trigger RNAi, such as small interfering RNA (siRNA) and plasmid DNA (pDNA) expressing short hairpin RNA (shRNA), present drawbacks arising from innate immune stimulation. To overcome them, an intelligent shRNA expression device (iRed) designed to induce RNAi was developed. The minimum sequence of iRed encodes only the U6 promoter and shRNA. A series of iRed comprises a polymerase chain reaction (PCR)-amplified 4′-thioDNA in which any one type of adenine (A), guanine (G), cytosine (C), or thymine (T) nucleotide unit was substituted by each cognate 4′-thio derivatives, i.e., dSA iRed, dSG iRed, dSC iRed, and ST iRed respectively. Each modified iRed acted as a template to transcribe shRNA with RNAi activity. The highest shRNA yield was generated using dSC iRed that exerted gene silencing activity in an orthotopic mouse model of mesothelioma. Reducing the minimal structure required to transcribe shRNA and the presence of the 4′-thiomodification synergistically function to abrogate innate immune response induced by dsDNA. The iRed will introduce a new approach to induce RNAi without inducing a detectable innate immune response.
Journal Title
Molecular Therapy - Nucleic Acids
ISSN
21622531
Publisher
nature
Volume
5
Start Page
e274
Sort Key
e274
Published Date
2016-01-05
Remark
© 2016 Tarashima et al. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article′s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http:// creativecommons.org/licenses/by/4.0/
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language
eng
TextVersion
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departments
Pharmaceutical Sciences