title: Single-Molecule Fluorescence Energy Transfer Assays for the Characterization of Reaction Pathways of miRNA-Argonaute Complex.
abstract: Argonaute proteins are key components of the microRNA-induced silencing complexes (miRISCs) that mediate the posttranscriptional gene silencing of microRNAs and small interfering RNA (siRNAs). The complex reaction mechanism of miRISC is expected to be characterized by tracing the reaction pathways of miRISC at the single-molecule level in real time. In this chapter, we describe single-molecule fluorescence resonance energy transfer (FRET) assays to observe the target binding and reaction pathways of miRISC composed of a recombinant Argonaute and a small RNA.
authors.affiliation: Department of Physics and Astronomy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea.|Institute of Applied Physics, Seoul National University, Seoul, 08826, Republic of Korea.|National Center of Creative Research Initiatives, Seoul National University, Seoul, 08826, Republic of Korea.
authors.forename: Myung Hyun
authors.initials: MH
authors.lastname: Jo
authors.affiliation: Department of Physics and Astronomy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea. shohng@snu.ac.kr.|Institute of Applied Physics, Seoul National University, Seoul, 08826, Republic of Korea. shohng@snu.ac.kr.|National Center of Creative Research Initiatives, Seoul National University, Seoul, 08826, Republic of Korea. shohng@snu.ac.kr.|Department of Biophysics and Chemical Biology, Seoul National University, Seoul, 08826, Republic of Korea. shohng@snu.ac.kr.
authors.forename: Sungchul
authors.initials: S
authors.lastname: Hohng
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000737:chemistry* / Q000235:genetics; D004330:Drosophila; D005453:Fluorescence; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D035683:MicroRNAs / Q000737:chemistry* / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics; D034743:RNA-Induced Silencing Complex / Q000737:chemistry / Q000235:genetics*
chemical_list: D060565:Argonaute Proteins; D035683:MicroRNAs; D034741:RNA, Small Interfering; D034743:RNA-Induced Silencing Complex
keywords: Argonaute; Single-molecule FRET; miRNA; siRNA