Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Quantitative Analysis of Mitochondrial RNA in Living Cells with a Dual-Color Imaging System
Small ( IF 13.0 ) Pub Date : 2023-05-01 , DOI: 10.1002/smll.202301132
Zhiyan Chen 1 , Su Zeng 1 , Linghui Qian 1
Small ( IF 13.0 ) Pub Date : 2023-05-01 , DOI: 10.1002/smll.202301132
Zhiyan Chen 1 , Su Zeng 1 , Linghui Qian 1
Affiliation
![]() |
Accurate quantification and dynamic expression profiling of mitochondrial RNA (mtRNA for short) are critical for illustrating their cellular functions. However, there lack methods for precise detection of mtRNA in situ due to the delivery restrictions and complicated cellular interferences. Herein, a dual-color imaging system featured with signal amplification and normalization capability for quantitative analysis of specific mtRNA is established. As a proof-of-concept example, an enzyme-free hairpin DNA cascade amplifier fine-tailored to specifically recognize mtRNA encoding NADH dehydrogenase subunit 6 (ND6) is employed as the signal output module and integrated into the biodegradable mitochondria-targeting black phosphorus nanosheet (BP-PEI-TPP) to monitor spatial-temporal dynamics of ND6 mtRNA. An internal reference module targeting β-actin mRNA is sent to the cytoplasm via BP-PEI for signal normalization, facilitating mtRNA quantification inside living cells with a degree of specificity and sensitivity as high as reverse transcription-quantitative polymerase chain reaction (RT-qPCR). With negligible cytotoxicity, this noninvasive “RT-qPCR mimic” can accurately indicate target mtRNA levels across different cells, providing a new strategy for precise analysis of subcellular RNAs in living systems.
中文翻译:
使用双色成像系统定量分析活细胞中的线粒体 RNA
线粒体 RNA(简称 mtRNA)的准确定量和动态表达谱对于阐明其细胞功能至关重要。然而,由于递送限制和复杂的细胞干扰,缺乏原位精确检测 mtRNA 的方法。在此,建立了一种具有信号放大和归一化能力的双色成像系统,用于特定 mtRNA 的定量分析。作为概念验证示例,采用经过精心设计的无酶发夹 DNA 级联放大器作为信号输出模块,专门识别编码 NADH 脱氢酶亚基 6 (ND6) 的 mtRNA,并将其集成到可生物降解的线粒体靶向黑磷纳米片中( BP-PEI-TPP ) 监测 ND6 mtRNA 的时空动态。针对 β-actin mRNA 的内部参考模块通过BP-PEI发送到细胞质进行信号标准化,促进活细胞内 mRNA 定量,其特异性和灵敏度与逆转录定量聚合酶链反应 (RT-qPCR) 一样高。这种非侵入性“RT-qPCR 模拟物”的细胞毒性可以忽略不计,可以准确指示不同细胞中的目标 mRNA 水平,为精确分析生命系统中的亚细胞 RNA 提供了一种新策略。
更新日期:2023-05-01
中文翻译:

使用双色成像系统定量分析活细胞中的线粒体 RNA
线粒体 RNA(简称 mtRNA)的准确定量和动态表达谱对于阐明其细胞功能至关重要。然而,由于递送限制和复杂的细胞干扰,缺乏原位精确检测 mtRNA 的方法。在此,建立了一种具有信号放大和归一化能力的双色成像系统,用于特定 mtRNA 的定量分析。作为概念验证示例,采用经过精心设计的无酶发夹 DNA 级联放大器作为信号输出模块,专门识别编码 NADH 脱氢酶亚基 6 (ND6) 的 mtRNA,并将其集成到可生物降解的线粒体靶向黑磷纳米片中( BP-PEI-TPP ) 监测 ND6 mtRNA 的时空动态。针对 β-actin mRNA 的内部参考模块通过BP-PEI发送到细胞质进行信号标准化,促进活细胞内 mRNA 定量,其特异性和灵敏度与逆转录定量聚合酶链反应 (RT-qPCR) 一样高。这种非侵入性“RT-qPCR 模拟物”的细胞毒性可以忽略不计,可以准确指示不同细胞中的目标 mRNA 水平,为精确分析生命系统中的亚细胞 RNA 提供了一种新策略。