The purpose of this proposal is to continue the development of our high resolution, sensitive FISH technology and apply it to the study of RNA movements in living cells in real time. 1. To extend the technology of detecting RNA transcripts in living cells to detect single molecules in living cells. 2. To describe the movements of RNA transcripts in the cytoplasm using methods capable of single molecule detection and distinguish among various models for RNA localization. 3. To use high speed microscopic 4D imaging to follow movements of RNA transcripts in the nucleus in vivo and develop algorithms to follow the movements of hundreds of molecules over a short time window to distinguish diffusional from directed events in the nucleus. This work will result in the ability to define mechanisms which regulate the spatial aspects of RNA metabolism: RNA nuclear export, cytoplasmic transport and localization, important for the normal functioning of all eucaryotic cells.
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| Ben-Ari, Ya'ara; Brody, Yehuda; Kinor, Noa et al. (2010) The life of an mRNA in space and time. J Cell Sci 123:1761-74 |
| Hocine, Sami; Singer, Robert H; Grünwald, David (2010) RNA processing and export. Cold Spring Harb Perspect Biol 2:a000752 |
| Wang, Donghai; Pezo, Rossanna C; Corner, Georgia et al. (2010) Altered dynamics of intestinal cell maturation in Apc1638N/+ mice. Cancer Res 70:5348-57 |
| Grünwald, David; Singer, Robert H (2010) In vivo imaging of labelled endogenous ?-actin mRNA during nucleocytoplasmic transport. Nature 467:604-7 |
| Park, Hye Yoon; Buxbaum, Adina R; Singer, Robert H (2010) Single mRNA tracking in live cells. Methods Enzymol 472:387-406 |
| Dib, Amel; Glebov, Oleg K; Shou, Yaping et al. (2009) A der(8)t(8;11) chromosome in the Karpas-620 myeloma cell line expresses only cyclin D1: yet both cyclin D1 and MYC are repositioned in close proximity to the 3'IGH enhancer. DNA Repair (Amst) 8:330-5 |
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| Rodriguez, Alexis J; Czaplinski, Kevin; Condeelis, John S et al. (2008) Mechanisms and cellular roles of local protein synthesis in mammalian cells. Curr Opin Cell Biol 20:144-9 |
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