Background

Transient expression of proteins in mammalian cells is a key technique for many functional and structural studies of human and higher eukaryotic genes as well as for the production of recombinant protein therapeutics. Maximizing the expression efficiency to achieve a higher expression yield is desirable and may be even critical when, for instance, an expressed protein must be characterized at the single-cell level.

New Methods

Our goal was to develop a simple method by which protein expression yield in human embryonic kidney (HEK)-293 cells could be enhanced with a brief treatment of dimethyl sulfoxide (DMSO) solution.

Results

By expressing green fluorescent protein (GFP) as a reporter protein using the calcium phosphate transfection method and imaging a large population of cells, we found that a 5-min exposure of 10 % DMSO to HEK-293 cells, 4 h after transfection of the protein of interest, leads to ∼1.6-fold increase in the expression yield without causing any appreciable cytotoxicity. By expressing an α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and separately a kainate receptor in HEK-293 cells and measuring glutamate-induced whole-cell current response, we also found that such a brief DMSO treatment did not affect channel activity.

Conclusion

This method is simple, efficient and inexpensive to use for enhancing transient transfection yield in HEK-293 cells.

中文翻译：

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通过将培养物短暂暴露于 DMSO 来增强 HEK-293 细胞中的瞬时蛋白质表达

背景

哺乳动物细胞中蛋白质的瞬时表达是人类和高等真核生物基因的许多功能和结构研究以及重组蛋白质治疗剂生产的关键技术。最大化表达效率以实现更高的表达产量是可取的，甚至可能是至关重要的，例如，当必须在单细胞水平上表征表达的蛋白质时。

新方法

我们的目标是开发一种简单的方法，通过该方法可以通过简单地处理二甲基亚砜 (DMSO) 溶液来提高人胚胎肾 (HEK)-293 细胞中的蛋白质表达产量。

结果

通过使用磷酸钙转染法表达绿色荧光蛋白 (GFP) 作为报告蛋白并对大量细胞进行成像，我们发现在转染 4 小时后，将 10% DMSO 暴露于 HEK-293 细胞 5 分钟。感兴趣的蛋白质，导致表达产量增加约 1.6 倍，而不会引起任何明显的细胞毒性。通过在 HEK-293 细胞中表达 α-氨基-3-羟基-5-甲基-4-异恶唑丙酸 (AMPA) 和单独的红藻氨酸受体并测量谷氨酸诱导的全细胞电流反应，我们还发现这样一个简短的DMSO 处理不影响通道活性。

结论

该方法简单、高效且成本低廉，可用于提高 HEK-293 细胞的瞬时转染产量。