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菜:通过基于光的选择和定向进化的快速选择,对绿色光蛋白的RNA模拟进行快速选择

Grigory S Filonov¹, Jared D Moon, Nina Svensen

¹Department of Pharmacology, Weill Cornell Medical College , New York, New York 10065, United States.

Journal of the American Chemical Society

|October 23, 2014

概括

研究人员开发了一种新方法来创建高度光的RNA分子用于活细胞成像. 这种方法快速生成新的RNA-光体复合体,比如菜的吸收体,用于改进细胞应用.

科学领域:

分子生物学分子生物学
生物化学生物化学
细胞生物学细胞生物学

背景情况:

基因编码的光核糖酸 (RNA) 对活细胞成像和基于RNA的传感器至关重要.
像菜和菜2这样的现有RNA吸收体激活光体的光,但在细胞环境中存在局限性.
鉴定新型,高度光的RNA-光体复合物是由于当前的选择方法具有挑战性的.

研究的目的:

开发一个平台,快速生成高光的RNA-光体复合体,优化为细胞功能.
与现有RNA相比,识别具有增强光和细胞性能的新型RNA体.

主要方法:

一个新的选择平台,结合了阿普坦酶与光激活细胞分类 (FACS) 结合在菌中.
利用基于FACS的定向进化来进一步优化有前途的aptamer候选者.
在活细胞中描述了新发现的阿普塔默的表现,例如西兰花阿普塔默.

主要成果:

确定了几种新型RNA吸收体,包括49核酸布洛克利吸收体.
花在细胞中表现出强大的折叠和绿色光,表现优于菜2,特别是在低的条件下.
新的选择方法简化了针对细胞表达和性能优化的体的生成.

结论:

开发的基于光的选择平台加速了功能性RNA-光体复合物的发现.
布洛克利阿普坦体代表了光RNA技术在活细胞应用中的重大进步.
这种方法克服了先前方法的局限性,使得能够有效地优化细胞环境中的aptamers.

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