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活细胞中的微RNA和端粒酶的双量定量

Wei Ma^1,2,3, Pan Fu^1,2,3, Maozhong Sun^1,2,3

¹State Key Lab of Food Science and Technology, Jiangnan University , Wuxi, Jiangsu 214122, People's Republic of China.

Journal of the American Chemical Society

|August 2, 2017

概括

研究人员开发了一种新的双信号平台,用于同时检测活细胞中的microRNA-21和端粒酶. 这种方法可实现对这些关键生物分子的超敏感量化,有助于诊断疾病.

科学领域:

生物医学工程
分析化学
分子生物学

背景情况:

同时对活细胞中的核酸和蛋白质等多种生物分子进行量化具有重大挑战.
现有的方法往往缺乏在现场细胞分析所需的灵敏度或通用性.

研究的目的:

开发一个通用和超灵敏的平台,同时在现场定量检测活细胞中的microRNA-21和端粒酶.
建立一个双信号策略,整合表面增强的拉曼散射 (SERS) 和上转换发光.

主要方法:

使用基于SERS的方法检测微RNA-21,其中与分子信标混合改变SERS信号强度.
通过酶活性和随后的纳米粒子释放来触发端粒酶检测的上转化发光策略.
使用拉曼和共聚焦成像验证了平台,显示出高灵敏度和特异性.

主要成果:

实现了细胞内微RNA-21的超敏感检测,检测极限为0. 011 amol/ ng_RNA.
量化细胞内端粒酶的LOD为3. 2×10−13IU,在特定的度范围内呈现线性反应.
在活细胞内同时精确检测两种分析物的能力.

结论:

开发的双信号平台为细胞系统中多个生物分子的同时和定量分析提供了强大的工具.
这种创新方法增强了对细胞化学的理解,并有可能用于疾病诊断.
该策略的灵敏性和通用性为复杂的生物环境中检测各种信号分子铺平了道路.

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相关概念视频

MicroRNAs

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...

MicroRNAs

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...