Digital PCR and intra-cellular expression profiling

Mikael Kubista 1 2, Monika Sidova 1,2, David Svec 1,2, Radek Sindelka 1,2
1 TATAA Biocenter; 2 Institute of Biotechnology, Czech Academy of Sciences

Expression profiling of single cells reveal substantial heterogeneity of unrelated mRNAs, while transcripts with related functions show correlated expression levels, evidencing stringent temporal control of genes’ expressions. In my talk I address the spatial distribution of mRNAs within a cell. The Xenopus laevis oocyte is excellent model for these studies, because of its large size, high content of mRNA, and differentially colored poles, which allow the cell to be aligned, mounted, and sliced. Measuring the mRNA levels in the slices with RT qPCR reveals three classes of mRNAs with distinct special distributions. The presence of these intra cellular mRNA gradients is further confirmed with digital PCR. Our finding suggests that before cell division mRNAs may become spatially distributed within the cells such that the progeny cells obtain different mRNA contents and thus become predisposed for differentiation. We postulate this is general mechanism behind asymmetric cell division.

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