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[14p-P9-8] Protein introduction into plant cells using a multi-gas plasma jet
Keywords:non-thermal atmospheric-pressure plasma, protein introduction, plant cells
Protein introduction into living cells is one of an important technique in not only basic research but also industrial uses. However, protein introduction into plant cells is more difficult than that into mammalian cells, because plants have a unique structure of cell surface and cell walls. So far, it was only reported that proteins were introduced into the cells of shoot apical meristem and leaves together with a cell-penetrating peptide in plants. However, their methods have restriction to use in plant spices and tissues. Thus, novel techniques are expected to be developed for using in various plant spices and tissues.
We tried to introduce sGFP (sperfolder green fluorescence protein) fusion protein without any cell-penetrating peptide into the cells of tobacco leaves by treatment with non-thermal atmospheric-pressure plasmas. For this purpose, a multi-gas plasma jet source was used (Plasma Concept Tokyo, PCT-DFPJ02). The body of the device was grounded and the interior high-voltage electrode was connected to an AC power supply of 16 kHz and 9 kV (pproximately 10 W). CO2 or N2 gas was used to generate a stable atmospheric-pressure plasma through a 1 mm hole with a flow rate of 5 L/min. For making lower temperature plasma (approximately 20-30 oC), the gas was cooled using a gas-cooling device that uses liquid nitrogen.
Confocal microscopy indicated that the fluorescence of sGFP was observed inside of leaf cells after treatment with CO2 or N2 plasma without substantial damage. However, no sGFP signal was observed in any sample treated by gas, which was a negative control. These results indicated that the treatment using the multi-gas plasma jet enabled direct protein introduction into plant cells.
We tried to introduce sGFP (sperfolder green fluorescence protein) fusion protein without any cell-penetrating peptide into the cells of tobacco leaves by treatment with non-thermal atmospheric-pressure plasmas. For this purpose, a multi-gas plasma jet source was used (Plasma Concept Tokyo, PCT-DFPJ02). The body of the device was grounded and the interior high-voltage electrode was connected to an AC power supply of 16 kHz and 9 kV (pproximately 10 W). CO2 or N2 gas was used to generate a stable atmospheric-pressure plasma through a 1 mm hole with a flow rate of 5 L/min. For making lower temperature plasma (approximately 20-30 oC), the gas was cooled using a gas-cooling device that uses liquid nitrogen.
Confocal microscopy indicated that the fluorescence of sGFP was observed inside of leaf cells after treatment with CO2 or N2 plasma without substantial damage. However, no sGFP signal was observed in any sample treated by gas, which was a negative control. These results indicated that the treatment using the multi-gas plasma jet enabled direct protein introduction into plant cells.