Book of Abstracts: Albany 2009
June 16-20 2009
© Adenine Press (2008)
Analysis of reprogramming of cellular fate induced by a large non-coding RNA
Large non-coding RNAs, are thought to form a vast and complex layer of regulatory mechanisms that have so far remained almost completely unknown. In order to gain insight into the function of this novel class of RNAs, we chose a large mRNA-like transcript that lacked protein-coding potential for analysis. We initially determined the expression pattern of this RNA in different tissues in mouse, which showed significant expression in neuroal tissues of both adult and fetal mouse, including forebrain, brainstem and cerebellum. The expression of this RNA in other tissues was either barely detectable or completely absent. Analysis of primary cultured cells derived from forebrain indicated that the RNA was highly expressed in neurons, but not in glial cells. Thus, the expression of the RNA was highly restricted to neurons, suggesting that it might play a role in neuronal differentiation or maintenance. Intriguingly, over-expression of this RNA in a number of cell lines, including the C2C12 mouse myoblasts and C3H10T1/2 fibroblasts, resulted in reprogramming of the differentiation pathway of these cells from muscle or adipocyte cells into neurons. In order to gain insight into the mechanism of this drastic cell fate switch, we have started to determine the point at which the myogenic or adipogenic programs are diverted into a neurogenic pathway. To this end, we have analyzed the gene expression pattern of stable cell lines that overexpress the noncoding RNA at different stages before and during differentiation. Our preliminary analysis suggests that while embryonic and neuronal stem cell markers are not expressed in these cells, genes upregulated in Neural Progenitor Cells and mature neurons are highly expressed in stable cell lines that overexpress the large RNA. This was not observed in control cells, or vector transfected cells. Experiments are underway to further characterize the mechanism of this cell fate switch. These data indicate that large non-coding RNAs can function as master regulatory molecules in cells, and underscore the critical role of RNAs in cellular function.
Center for RNA Molecular Biology