Development of Novel Small Hairpin RNAs That do not Require Processing by Dicer or AGO2.
The innate cytokine response to nucleic acid is the most challenging problem confronting the practical use of nucleic acid medicine. The degree of stimulation of the innate cytokine response strongly depends on the length of the nucleic acid. In this study, we developed a 30-nucleotide single-strand RNA, termed "guide hairpin RNA (ghRNA, ghR)", that has a physiological function similar to that of miRNA and siRNA. The ghR caused no innate cytokine response either in vitro or in vivo. In addition, its structure does not contain a passenger strand seed sequence, reducing the unwanted gene repression relative to existing short RNA reagents. Systemic and local injection of ghR-form miR-34a (ghR-34a) suppressed tumor growth in a mouse model of RAS-induced lung cancer. Furthermore, Dicer and AGO2 are not required for ghR-34a function. This novel RNA interference (RNAi) technology may provide a novel, safe, and effective nucleic acid drug platform that will increase the clinical usefulness of nucleic acid therapy.
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Shin-Ichiro
S
Ohno
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Karen
K
Itano
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Yuichirou
Y
Harada
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Koutaro
K
Asada
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Keiki
K
Oikawa
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Mikie
M
Kashiwazako
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Hikaru
H
Okuyama
Department of Animal Research Center, Tokyo Medical University, Tokyo, Japan.
Katsuyoshi
K
Kumagai
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Masakatsu
M
Takanashi
Department of Animal Research Center, Tokyo Medical University, Tokyo, Japan.
Katsuko
K
Sudo
Department of Respiratory Surgery, Tokyo Medical University, Tokyo, Japan.
Norihiko
N
Ikeda
Department of Molecular Pathology, Tokyo Medical University, Tokyo, Japan.
Masahiko
M
Kuroda
D000818:Animals; D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism*; D020029:Base Pairing; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002460:Cell Line; D004195:Disease Models, Animal; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology; D051379:Mice; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism*; D000094704:RNA, Guide, CRISPR-Cas Systems
D060565:Argonaute Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D043244:Ribonuclease III; D000094704:RNA, Guide, CRISPR-Cas Systems; C466422:AGO2 protein, human