Abstract
Live cell imaging has been instrumental in analyzing the dynamic properties of RNA. New technologies in optical microscopy and fluorescent probe development have been pushing the envelope of our analysis capabilities. We have been dedicated to developing and implementing these technologies to further the understanding of single mRNA dynamics in cells and organisms. We have utilized computational approaches to analyze real-time transcription activities of endogenous genes from yeast to human cells. We have employed a plethora of imaging methods, ranging from confocal and multiphoton microscopy, long-term cell imaging, high-speed real-time widefield microscopy, single molecule tracking, and we have developed super-registration microscopy and fluorescence fluctuation analysis. We have investigated key processes of RNA synthesis: initiation, elongation, termination, as well as nuclear pore export, cytoplasmic trafficking, localization and decay. Mathematical modeling allowed us to extract quantitative kinetic parameters that precisely describe these processes in living cells.Supported by NIH GMS Grants to RH Singer.
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