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Séminaire impromptu - Dan Othan WANGSynaptic epitranscriptomics and dynamic RNA imaging

Abstract :

 One of the main challenges in the post-genomic era is to decipher the dynamic spatiotemporal organization of gene expression in relation to cell function.
This is exemplified by the versatile cis- and trans-, protein- and RNA-mediated regulation identified in every known aspect of life. To decipher the dynamics of RNA expression profiles in space and time, we have developed new quenching-based RNA imaging techniques. Our imaging methods allow endogenous RNA to be labeled in intact animals and imaged in living tissues, opening the possibility to study RNA dynamics in their physiological context. Already we have applied our imaging techniques to test RNA foci dynamics and phase-transition physical models in living cells.

In addition to pioneering novel imaging techniques, our lab seeks to elucidate unknown mechanisms cells use to regulate gene expression in neurons. Methyl-N6-adenosine (m6A) has recently been identified in thousands of mRNAs in embryonic and adult brains, offering additional complexity in post-transcriptional regulation. We developed an m6A-seq protocol using sub-micromolar quantities of input RNA, enabling characterization of an m6A-epitranscriptome in adult mouse forebrain synaptosomes. We found m6A peaks enriched in diverse synaptic RNA species independent of synaptic enrichment of the cognate gene, indicating these transcripts may be preferentially methylated at synapses.
Gene-annotation enrichment analysis indicates functional clustering in various cellular processes including neural development and growth. These results indicate synaptic function may be regulated through dynamic epitranscriptomic marks of the local transcriptome at the synapse.

Selected publications

ECHO-liveFISH: in vivo RNA labeling reveals dynamic regulation of nuclear RNA foci in living tissues.
Oomoto I, Suzuki-Hirano A, Umeshima H, Han YW, Yanagisawa H, Carlton P, Harada Y, Kengaku M, Okamoto A, Shimogori T, Wang DO. Nucleic Acids Res. 2015 Oct 30;43(19):e126. doi: 10.1093/nar/gkv614. Epub 2015 Jun 22.

Visualization of nucleic acids with synthetic exciton-controlled fluorescent oligonucleotide probes.
Wang DO, Okamoto A. Methods Mol Biol. 2015;1262:69-87. doi: 10.1007/978-1-4939-2253-6_5.


Scientific focus :

Dan Ohtan WANG
Wang graduated in 2004 with a Ph.D. degree from University of Southern California after studying neuroscience under Dr. Michael W. Quick. She became a chief researcher at Kyoto University’s WPI-iCeMS Program in 2012. Wang is interested in the gene expression that results in neural plasticity, a fundamental aspect of molecular and cellular learning mechanisms.