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Spatially Selective Imaging of Mitochondrial MicroRNAs via Optically Programmable Strand Displacement Reactions
Jian Zhao1,2, Zhixiang Li1,3, Yulei Shao1,2
1CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety and CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology, Beijing, 100190, China.
Abstract:
MicroRNA (miRNA) functions are tightly regulated by their sub-compartmental location in living cells, and the ability to imaging of mitochondrial miRNAs (mitomiRs) is essential for understanding of the related pathological processes. However, most existing DNA-based methods could not be used for this purpose. Here, we report the development of a DNA nanoreporter technology for imaging of mitomiRs in living cells through near-infrared (NIR) light-controlled DNA strand displacement reactions. The sensing function of the DNA nanoreporters are silent (OFF) during the delivery process, but can be photoactivated (ON) with NIR light after targeted mitochondrial localization, enabling spatially-restricted imaging of two types of cancer-related mitomiRs with improved detection accuracy. Furthermore, we demonstrate imaging of mitomiRs in vivo through spatiotemporally-controlled delivery and activation. Therefore, this study illustrates a simple methodology that may be broadly applicable for investigating the mitomiRs-associated physiological events.
Insights
Researchers developed novel DNA nanoreporters for imaging mitochondrial microRNAs (mitomiRs) in living cells. This near-infrared light-activated technology enables precise, spatially-controlled detection of cancer-related mitomiRs in vitro and in vivo.
Area of Science:
- Molecular Biology
- Cell Biology
- Biotechnology
Background:
- MicroRNA (miRNA) localization within cellular compartments, particularly mitochondria, is crucial for their function and associated pathologies.
- Current imaging techniques are insufficient for visualizing mitochondrial miRNAs (mitomiRs) in living systems.
- Accurate imaging of mitomiRs is essential for understanding their role in disease.
Purpose of the Study:
- To develop a novel DNA nanoreporter technology for the specific imaging of mitomiRs in living cells.
- To enable photoactivated, spatially-restricted detection of mitomiRs using near-infrared (NIR) light.
- To demonstrate the in vivo applicability of the developed imaging technology.
Main Methods:
- Development of DNA nanoreporters utilizing NIR light-controlled DNA strand displacement reactions.
- Designing nanoreporters with a silent (OFF) sensing state during delivery and photoactivatable (ON) state post-mitochondrial localization.
- Testing the nanoreporters for imaging cancer-related mitomiRs in living cells and subsequently in vivo.
Main Results:
- Successful development of DNA nanoreporter technology for mitomiR imaging.
- Achieved photoactivation of sensing using NIR light after targeted mitochondrial delivery.
- Demonstrated spatially-restricted and accurate imaging of two cancer-related mitomiRs in living cells.
- Validated in vivo imaging of mitomiRs through spatiotemporally-controlled delivery and activation.
Conclusions:
- The developed DNA nanoreporter technology provides a powerful tool for imaging mitomiRs in living cells and organisms.
- This methodology offers a simple and broadly applicable approach for investigating mitomiR-associated physiological and pathological events.
- The photoactivatable nature allows for precise spatial and temporal control over mitomiR detection.
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