Studying Single mRNA Molecules from Cradle to Grave
We seek to understand the expression and movement of mRNA from transcription through degradation and the effect that defects in these processes have on health. We develop methods to label RNA in fixed and living cells using fluorescent probes and develop microscopy techniques and image analysis algorithms to visualize and quantify many mRNAs simultaneously. Using these technologies, we can observe single mRNAs localizing to cytoplasmic compartments such as the leading edge of a fibroblast, the bud tip of yeast or the axonal process of neurons and follow these single mRNAs from transcription, nuclear export through translation and degradation. Because these techniques yield quantitative fluorescence data, we are able to apply mathematical modeling to test mechanistic hypotheses.
Publications
- Visualization of Early RNA Replication Kinetics of SARS-CoV-2 by Using Single Molecule RNA-FISH Combined with Immunofluorescence.
Pathak R, Eliscovich C, Mena I, Cupic A, Rutkowska M, Chandran K, Jangra RK, García-Sastre A, Singer RH, Kalpana GV.
Viruses. 2024 Feb 7;16(2):262. doi: 10.3390/v16020262.PMID: 38400039 - Real-time single-molecule imaging of transcriptional regulatory networks in living cells.
Hwang DW, Maekiniemi A, Singer RH, Sato H.
Nat Rev Genet. 2024 Apr;25(4):272-285. doi: 10.1038/s41576-023-00684-9. Epub 2024 Jan 9.PMID: 38195868 - RNA and Protein Detection by Single-Molecule Fluorescent in Situ Hybridization (smFISH) Combined with Immunofluorescence in the Budding Yeast S. cerevisiae.
Maekiniemi A, Singer RH.
Methods Mol Biol. 2024;2784:45-58. doi: 10.1007/978-1-0716-3766-1_3.PMID: 38502477 - Michael R. Green (1954-2023).
Roeder RG, Deibler SK, Jacobson A, Lillie JW, Rosbash M, Singer RH, Wajapeyee N
.Mol Cell. 2023 Jul 6;83(13):2152-2154. doi: 10.1016/j.molcel.2023.06.027.PMID: 37419086 - Maintenance of a short-lived protein required for long-term memory involves cycles of transcription and local translation.
Das S, Lituma PJ, Castillo PE, Singer RH.
Neuron. 2023 Jul 5;111(13):2051-2064.e6. doi: 10.1016/j.neuron.2023.04.005. Epub 2023 Apr 25.PMID: 37100055