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在动物细胞中诱导性转基因表达的RNA Aptamer介导的基因激活系统

Feiyang Zheng¹, Yoshinori Kawabe¹, Masamichi Kamihira¹

¹Department of Chemical Engineering, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.

ACS synthetic biology

|December 11, 2023

概括

研究人员开发了一种新型的RNA胺酶介导基因激活 (RAMGA) 系统. 这种合成生物学工具可以实时监测和调制活细胞内的RNA表达,增强基因电路开发和RNA检测能力.

科学领域:

合成生物学合成生物学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

RNA表达分析为细胞状态提供了洞察力.
实时细胞内RNA监测对于理解基因调节至关重要.
合成生物学为精确的RNA检测和分析提供了工具.

研究的目的:

开发一种RNA胺酶介导的基因激活 (RAMGA) 系统.
为了使RNA触发基因表达激活,使用合成生物学原理.
为实时细胞内RNA检测和调制创建一个平台.

主要方法:

设计了一个可诱导的复合形成策略,使用RNA结合域 (MS2,PP7) 和事务激活器.
融合的MS2菌体外套蛋白与转录激活剂和PP7外套蛋白与四环素抑制剂 (tetR).
生成的重组CHO细胞具有可诱导的GFP表达模块,由R响应元素调节.

主要成果:

具有触发RNA的细胞表现出强大的记者基因 (GFP) 表达,与对照细胞相比增加了200倍以上.
没有触发RNA的细胞没有显著的记者基因表达.
该系统成功检测了用aptamer标签标记的mRNA,并根据目标mRNA水平调节了基于目标mRNA的记者基因表达,而不会影响原始mRNA.

结论:

拉姆加系统为RNA触发的基因表达提供了一个强大的平台.
这项技术有助于评估内源基因表达和开发新型RNA检测器.
开发的RNA触发基因表达系统有可能建立复杂的基因电路.

关键词:

这是一种RNA合酶.检测RNA检测RNA的检测基因表达系统基因表达系统转录激活的转录激活

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