title: Simultaneous live imaging of the transcription and nuclear position of specific genes.
abstract: The relationship between genome organization and gene expression has recently been established. However, the relationships between spatial organization, dynamics, and transcriptional regulation of the genome remain unknown. In this study, we developed a live-imaging method for simultaneous measurements of the transcriptional activity and nuclear position of endogenous genes, which we termed the 'Real-time Observation of Localization and EXpression (ROLEX)' system. We demonstrated that ROLEX is highly specific and does not affect the expression level of the target gene. ROLEX enabled detection of sub-genome-wide mobility changes that depended on the state of Nanog transactivation in embryonic stem cells. We believe that the ROLEX system will become a powerful tool for exploring the relationship between transcription and nuclear dynamics in living cells.
authors.affiliation: Research Center for the Mathematics on Chromatin Live Dynamics (RcMcD), Hiroshima University, Higashi-Hiroshima 739-8530, Japan ochiai@hiroshima-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ochiai
authors.affiliation: Research Center for the Mathematics on Chromatin Live Dynamics (RcMcD), Hiroshima University, Higashi-Hiroshima 739-8530, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sugawara
authors.affiliation: Research Center for the Mathematics on Chromatin Live Dynamics (RcMcD), Hiroshima University, Higashi-Hiroshima 739-8530, Japan Department of Mathematical and Life Sciences, Graduate School of Science, Hiroshima University, Higashi-Hiroshima 739-8526, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics; D002467:Cell Nucleus / Q000235:genetics*; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000378:metabolism; D004720:Endonucleases / Q000032:analysis / Q000235:genetics; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D008810:Mice, Inbred C57BL; D000071317:Nanog Homeobox Protein; D061848:Optical Imaging / Q000379:methods; D039904:Pluripotent Stem Cells / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis; D014158:Transcription, Genetic
chemical_list: D036022:Capsid Proteins; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004720:Endonucleases