An enhanced platform for cell electroporation: controlled delivery and electrodes functionalization

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Functionalized microelectrodes arrays with integrated microfluidic channels for single-site multiple

Nowadays, different chemical and physical transfection techniques are used to delivery biomolecules of interest (e.g. DNA, RNA, proteins) into cells. Among the physical methods, electroporation generates transient pores in the plasma membrane by applying electrical pulses to suspended cells. One of its main limitations is the lack of spatio-temporal control over the process: it does not allow to select single cells (desirable requirement especially in highly heterogeneous tissues), and to monitor the transfection results in real-time. To circumvent these disadvantages, alternative microscale approaches are increasingly required. This work presents an integrated platform consisting of a gold microelectrode array (MEA) for singlesite electroporation and fluidic channels for controlled delivery of bio-chemical entities . In order to improve the efficiency of electroporation, the gold electrodes were coated with a thin film of nanostructured of Titanium Dioxide

Intracellular trafficking of RNASET2, a novel component of P-bodies

Here, we have studied the intracellular trafficking of RNASET2 in mammalian cells. RNASET2 co-localizes with markers for the trans-Golgi network (TGN), which is the central sorting and processing station of the secretory pathway. Moreover, using the temperature-sensitive vesicular stomatitis glycoprotein, we demonstrate that RNASET2 undergoes delivery to the plasma membrane. In contrast to other RNA-interacting proteins, RNASET2 does not accumulate in stress granules upon metabolic stress in mammalian cells. Surprisingly, RNASET2 shows co-localization with processing bodies (P-bodies), which increases upon metabolic stress. Finally, cells lacking RNASET2 show a reduced numbers of P-bodies

An integrated platform for in vitro single-site cell electroporation: Controlled delivery and electrodes functionalization

This work presents an improved platform for single-site electroporation and controlled transfectants delivery. The device consists of a gold microelectrode array (MEA) with integrated microfluidics and nanostructured titanium dioxide (ns-TiO2) functionalized electrodes for the improvement of cell adhesion. Human cervical cancer cells (HeLa) have been successfully cultivated on chip surface using traditional protocols. The system has been previously tested by electroporating HeLa cells with Lucifer Yellow (LY) and then, in order to validate the approach and cell viability, with plasmid for the enhanced expression of Green Fluorescence Protein (pEGFP-N1) delivered through microchannels