Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P. O. BOX 1417863171, Tehran, Iran.
Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), P. O. BOX 1417863171, Tehran, Iran.
Ultrason Sonochem. 2021 May;73:105507. doi: 10.1016/j.ultsonch.2021.105507. Epub 2021 Mar 2.
Sonoporation, ultrasound-mediated membrane perforation can potentially puncture plasma membrane and rigid cell wall on presumably reversible basis which benefit gene transfection and plant biotechnology. Herein, positively charged poly-ethyleneimine (PEI)-coated mesoporous silica nanoparticles (MSNs) with an average diameter of 100 ± 8.7 nm was synthesized for GUS-encoding plasmid delivery into the suspended tobacco cells using the ultrasound treatment. The overall potential of PEI-MSN for DNA adsorption was measured at 43.43 μg DNA mg PEI-MSNs. It was shown that high level of sonoporation may adversely upset the cell viability. Optimal conditions of ultrasonic treatment are obtained as 8 min at 3 various intensities of 160, 320 and 640 W. Histochemical staining assay was used to follow the protein expression. It was shown that PEI-coated MSNs efficiently transfer the GUS-encoding plasmid DNA into the tobacco cells. The results of this study showed that ultrasonic treatment provides an economical and straightforward approach for gene transferring into the plant cells without any need to complicated devices and concerns about safety issues.
声孔作用，即超声介导的细胞膜穿孔，在可能的可逆基础上可以穿透质膜和刚性细胞壁，有利于基因转染和植物生物技术。本文中，我们合成了平均粒径为 100±8.7nm 的带正电的聚乙烯亚胺（PEI）包覆的介孔硅纳米颗粒（MSNs），并通过超声处理将携带 GUS 编码质粒的 MSNs 递送至悬浮烟草细胞。PEI-MSN 对 DNA 吸附的总潜能在 43.43μg DNA mg PEI-MSNs 时达到。研究表明，高强度的声孔作用可能会对细胞活力产生不利影响。在 3 种不同强度（160、320 和 640W）的超声处理 8 分钟可获得最佳条件。组织化学染色法用于跟踪蛋白表达。结果表明，PEI 包覆的 MSNs 可有效地将 GUS 编码质粒 DNA 转染至烟草细胞中。本研究结果表明，超声处理为基因转入植物细胞提供了一种经济、简便的方法，无需复杂的设备，也无需担心安全问题。
