title: Synthesis of native-like crosslinked duplex RNA and study of its properties.
abstract: A variety of enzymes have been found to interact with double-stranded RNA (dsRNA) in order to carry out its functions. We have endeavored to prepare the covalently crosslinked native-like duplex RNA, which could be useful for biochemical studies and RNA nanotechnology. In this study, the interstrand covalently linked duplex RNA was formed by a crosslinking reaction between vinylpurine (VP) and the target cytosine or uracil in RNA. We measured melting temperatures and CD spectra to identify the properties of the VP crosslinked duplex RNA. The crosslinking formation increased the thermodynamic stability without disturbing the natural conformation of dsRNA. In addition, a competitive binding experiment with the duplex RNA binding enzyme, ADAR2, showed the crosslinked dsRNA bound the protein with nearly the same binding affinity as the natural dsRNA, confirming that it has finely preserved the natural traits of duplex RNA.
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi 980-8577, Japan.
authors.forename: Kazumitsu
authors.initials: K
authors.lastname: Onizuka
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi 980-8577, Japan.
authors.forename: Madoka E
authors.initials: ME
authors.lastname: Hazemi
authors.affiliation: Department of Chemistry, University of California, Davis, One Shields Ave, Davis, CA 95616, USA.
authors.forename: Justin M
authors.initials: JM
authors.lastname: Thomas
authors.affiliation: Department of Chemistry, University of California, Davis, One Shields Ave, Davis, CA 95616, USA.
authors.forename: Leanna R
authors.initials: LR
authors.lastname: Monteleone
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi 980-8577, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Yamada
authors.affiliation: Faculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Imoto
authors.affiliation: Department of Chemistry, University of California, Davis, One Shields Ave, Davis, CA 95616, USA.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Beal
authors.affiliation: Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 2-1-1 Katahira, Aoba-ku, Sendai, Miyagi 980-8577, Japan. Electronic address: nagatugi@tagen.tohoku.ac.jp.
authors.forename: Fumi
authors.initials: F
authors.lastname: Nagatsugi
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D009690:Nucleic Acid Conformation; D012330:RNA, Double-Stranded / Q000138:chemical synthesis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D012330:RNA, Double-Stranded
keywords: 6-Vinylpurine; ADAR; Crosslinking reaction; Native-like; RNA