Background and Objectives mRNA-based protein expression technology has been used to express functional proteins. We have previously generated dopamine neurons from rat-embryo derived neural precursor cells (NPCs) through repeated transfection of synthetic transcription factor mRNA encoding dopamine-inducible genes. However, NPCs began to die approximately 10 d post-transfection. In this study, we examined a long-term transfection protocol that did not affect cell viability. Methods and Results Experiments were performed in eight groups sorted according to the start date of mRNA transfection. mRNA was transfected into NPCs daily for 21 d and live cell images of each group were recorded. NPCs which were differentiated for more than five days showed sustained gene expression and appreciable viability despite daily mRNA transfection for 21 d. Conclusions Repeated mRNA transfection requires cells with a sufficient differentiation period.

中文翻译：

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在分化的神经元中通过重复 mRNA 转染降低细胞毒性。

背景和目的 基于 mRNA 的蛋白表达技术已被用于表达功能蛋白。我们之前通过重复转染编码多巴胺诱导基因的合成转录因子 mRNA，从大鼠胚胎衍生的神经前体细胞 (NPC) 生成了多巴胺神经元。然而，NPC 在转染后约 10 天开始死亡。在这项研究中，我们检查了一种不影响细胞活力的长期转染方案。方法和结果 根据 mRNA 转染的开始日期，分八组进行实验。每天将 mRNA 转染到 NPC 中，持续 21 天，并记录每组的活细胞图像。尽管每天进行 mRNA 转染 21 天，分化超过五天的 NPC 仍显示出持续的基因表达和可观的生存能力。