一个新的,完全自动化的,与反应剂无关的短暂传染协议
Vasudha S Nair1, Kinnari Watson1
1Opentrons Labworks Inc, New York.
Current protocols
|February 5, 2024
概括
使用开源液体处理器进行过渡转染的新自动化协议在HeLa和A549细胞中实现了高效率. 这种方法可靠地引入核酸用于各种研究应用,包括基因治疗研究.
科学领域:
- 细胞生物学 细胞生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 转染对于将外来核酸引入真核细胞至关重要.
- 它可以研究细胞过程,疾病机制和基因疗法.
- 开发可靠和自动化的协议对于可重复性研究至关重要.
研究的目的:
- 提供可靠和完全自动化的过渡转染协议.
- 通过使用开源液体处理器和HEPA模块来评估传输效率.
- 为了比较不同细胞系和试剂的转化效率.
主要方法:
- 使用开源液体处理器和HEPA模块进行自动细胞播种和转染.
- 使用两个常见的转染试剂 (Lipofectamine 3000和FuGENE HD) 与GFP等离子体.
- 进行了图像采集和半定量分析,以确定HeLa和A549细胞的转染疗效.
主要成果:
- 在HeLa细胞中实现了从40.92%到73.26%的转染效率.
- 在难以转移的A549细胞系中获得了42.15%至54%的转化效率.
- 自动化协议的效率与手动方法相比相当.
结论:
- 开发的自动化协议为暂时转染提供了一种可靠的方法.
- 这种技术对标准细胞系和具有挑战性的细胞系都有效.
- 自动化提高了基因操纵研究的可复制性和效率.
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