Xi Zhao, Xiaomeng Huang, Xinmei Wang, Yun Wu, Ann-Kathrin Eisfeld, Sebastian Schwind, Daniel Gallego-Perez, Pouyan E. Boukany, Guido I. Marcucci and Ly James Lee
A comprehensive micro/nanofluidics platform for single-cell analysis based on nanochannel electroporation (NEP) and molecular beacon (MB) is presented in this study. The platform can quantitatively analyze multiple RNA species in individual cells with minimal cell damage. Furthermore, it is capable of delivering nucleic acids into target cells and subsequently detecting their responses at RNA level, e.g., microRNA (miRNA). It is known that as the downstream targets of miR-29b, DNMT3A/B can be downregulated by miR-29b overexpression. To demonstrate the activity of delivered miR-29b by NEP and the analytical function of the platform, the decreased expression of DNMT3A/B in acute myeloid leukemia (AML) cells was verified at single-cell level by simultaneous detection of multiple genes in the same cell. The potential of such platform on intracellular pathway studies has also been explored by investigating the upregulation efficiencies of miR-181a through different pathways in AML cells. The results showed that an indirect approach by C/EBPα-p30 peptide expression would have a stronger effect than direct transfection of the miR-181a gene. The platform has also shown its advantages over established technologies in the analysis of cells that are hard to transfect.
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