Seeing is believing: visualizing transcriptional dynamics in single cells.
For a gene to be expressed, the functions of multiple molecular machines must be coordinated at the site of transcription. To understand the role of nuclear organization in transcription, it is necessary to visualize the dynamic interactions of regulatory factors with chromatin and RNA. It is currently possible to localize individual transcription sites in single living mammalian cells by engineering reporter gene constructs to include sequence elements which permit the visualization of nucleic acids in vivo. Upon stable integration, these transgenes form chromatinized arrays, which can be imaged during activation to obtain high-resolution quantitative information about transcriptional dynamics. Modeling can suggest new hypotheses about gene regulation, which can be tested both in the single-cell imaging system and at endogenous genes. This gene-specific imaging strategy has the potential to reveal regulatory mechanisms, which would be difficult to imagine outside of single living cells.
Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania.
Alyshia
A
Newhart
Susan M
SM
Janicki
D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism*; D058948:Cell Tracking / Q000379:methods*; D002478:Cells, Cultured; D002843:Chromatin / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D004247:DNA / Q000378:metabolism*; D017930:Genes, Reporter; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008856:Microscopy, Fluorescence; D012313:RNA / Q000096:biosynthesis*; D014158:Transcription, Genetic; D014162:Transfection
D002843:Chromatin; D008164:Luminescent Proteins; D012313:RNA; D004247:DNA