首页|A Cell-Anchored and Self-Calibrated DNA Nanoplatform for in Situ Imaging and Quantification of Endogenous MicroRNA in Live Cells:Introducing Two Controls to Normalize the Sensing Signals

A Cell-Anchored and Self-Calibrated DNA Nanoplatform for in Situ Imaging and Quantification of Endogenous MicroRNA in Live Cells:Introducing Two Controls to Normalize the Sensing Signals

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Quantifying the microRNAs(miRNAs)levels in living cells,while essential for the study of fundamental biology and medical diagnostics,has barely been achieved due to insufficient probe delivery and unquantifiable signals.We report a cell-anchored and self-calibrated DNA nano-platform,a cholesterol-headed DNA nanowire that is capable of efficiently delivering to various cells and si-multaneously detecting two target miRNAs.One miRNA target can be utilized as an endogenous control against cell-to-cell variations.Moreover,the photocleavable lin-kers inserted in the nanostructures allow us to precisely regulate the probe structure and fluorescence signaling at the desired time and location in vivo.As a second control,the maximum fluorescence can be elicited by UV light,which further facilitates the normalization of the absolute fluorescence signal.With two introduced inter-nal controls,the maximum fluorescence and endogenous control gene,this approach displays excellent stability and self-calibration performance,effectively avoiding the interference from operating conditions and cell-to-cell variations,such as the laser powers and intracel-lular probe concentrations.Importantly,this design is capable of unifying the output signal intensity between in vitro test and cell imaging,making the in vitro linear calibration curve appropriate for the quantification of miRNA expression in living cells.

DNA self-assemblyquantification of miRNAfluorescence imaginginternal controlself-calibrationmembrane-anchoring groupphotoclea-vable probe

Wenjuan Song、Zhi-Ling Song、Qian Li、Chengwen Shang、Qiqi Chao、Xingfu Liu、Rongmei Kong、Gao-Chao Fan、Xiliang Luo

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Key Laboratory of Optic-Electric Sensing and Analytical Chemistry for Life Science,MOE,Shandong Key Laboratory of Biochemical Analysis,Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong,College of Chemistry and Molecular Engineering,Qingdao University of Science and Technology,Qingdao 266042

School of Chemistry and Chemical Engineering,Qufu Normal University,Qufu,Shandong 273165

Project Fund for Shangdong Key R&D Program国家自然科学基金Taishan Scholar Program of Shandong Province of ChinaState Key Laboratory of Analytical Chemistry for Life Science山东省自然科学基金Open Project of Chemistry Department of Qingdao University of Science and Technology

2019GSF10816921605091ts20110829SKLACLS2008ZR2021YQ11QUSTHX201926

2023

10.31635/ccschem.022.202101618

中国化学会会刊(英文)

中国化学会会刊(英文)

CSCD
ISSN：
年,卷(期)：2023.5(1)
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