title: Continuous Monitoring of Specific mRNA Expression Responses with a Fluorescence Resonance Energy Transfer-Based DNA Nano-tweezer Technique That Does Not Require Gene Recombination.
abstract: This letter discusses the feasibility of continuously monitoring specific mRNA expression responses in a living cell with a probe structured as a fluorescence resonance energy transfer (FRET)-based DNA nano-tweezer (DNA-NT). The FRET-based DNA-NT, self-assembled from three single-stranded DNAs, alters its structure from an open state to a closed state in recognition of a target mRNA, resulting in the closing of the distal relation of previously modified FRET-paired fluorescent dyes and generating a FRET signal. The expressions of glucose transporters (GLUT) 1 and 4 in a mouse hepato-carcinoma (Hepa 1-6 cells) were selected as the target model. Live-cell imaging analysis of Hepa 1-6 cells with both FRET-based DNA-NTs indicated that the behaviors of the FRET signals integrated in each individual cell were similar to those measured with the conventional mass analysis technique of semiquantitative real-time (RT) polymerase chain reaction (PCR). From these results, it is concluded that continuous monitoring of gene expression response without gene recombination is feasible with a FRET-based DNA-NT, even in a single cell manner.
authors.affiliation: Institute for Sustainable Sciences and Development, Hiroshima University , Higashihiroshima, Hiroshima 739-8511, Japan.|Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University , Higashihiroshima, Hiroshima 739-8530, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Shigeto
authors.affiliation: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University , Higashihiroshima, Hiroshima 739-8530, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Nakatsuka
authors.affiliation: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University , Higashihiroshima, Hiroshima 739-8530, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ikeda
authors.affiliation: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University , Higashihiroshima, Hiroshima 739-8530, Japan.
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Hirota
authors.affiliation: Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University , Higashihiroshima, Hiroshima 739-8530, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Kuroda
authors.affiliation: Institute for Sustainable Sciences and Development, Hiroshima University , Higashihiroshima, Hiroshima 739-8511, Japan.
authors.forename: Hisakage
authors.initials: H
authors.lastname: Funabashi
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D051272:Glucose Transporter Type 1 / Q000032:analysis* / Q000235:genetics; D051275:Glucose Transporter Type 4 / Q000032:analysis* / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000235:genetics*
chemical_list: D005456:Fluorescent Dyes; D051272:Glucose Transporter Type 1; D051275:Glucose Transporter Type 4; D012333:RNA, Messenger; C495678:Slc2a1 protein, mouse; C495693:Slc2a4 protein, mouse; D004247:DNA