Dual Energy Transfer-Based Fluorescent Nanoprobe for Imaging miR-21 in Nonalcoholic Fatty Liver Cells with Low Background

Chai, Shui Qin; Lv, Wen; He, Jia; Li, Chun Hong; Li, Yuan Fang; Li, Chun Mei; Huang, Cheng

doi:10.1021/acs.analchem.9b00841

Cited by 32 publications

(15 citation statements)

References 41 publications

(55 reference statements)

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“…Traditional single energy transfer nanoprobes are usually subject to limitations such as high background signals due to the low efficiency of resonance energy transfer (RET) between the donor and acceptor . In previous work, our group developed a dual energy transfer fluorescent nanoprobe containing a donor and two receptors by assembling gold nanoparticles (AuNPs) coupled with BHQ2 (AuBHQ) and quantum dots (QDs) that exhibited low background during imaging of miR-21 in nonalcoholic fatty liver cells . GO is typically used as an energy acceptor of various RET probes following long-range resonance energy transfer (LrRET). − BHQ was also chosen as the fluorescence resonance energy transfer (FRET) acceptor because it effectively quenches the fluorescence of the donor .…”

mentioning

confidence: 99%

Dual Energy Transfer-Based DNA/Graphene Oxide Nanocomplex Probe for Highly Robust and Accurate Monitoring of Apoptosis-Related microRNAs

et al. 2020

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Fluorescent labeled single-stranded DNA (ssDNA) molecules physisorbed on graphene oxide (GO) have been extensively explored as a useful sensing platform. However, this approach faces challenges when applied to complex biological samples due to heavy nonspecific desorption of nontarget molecules from GO. To overcome this problem, we introduced a capture DNA (cDNA) fragment with a poly adenine (poly-A) extension into the physisorption system that greatly reduces nonspecific desorption and false positive signal due to strong binding between poly-A and GO. Fluorescence from the dye can be effectively quenched by BHQ, which thus provides a second guarantee of anti-interference to avoid possible nonspecific poly-A DNA displacement. As a proof of concept, we have successfully developed a novel DNA-adsorbing GO nanocomplex probe (DNA-GO nanocomplex probe). This probe has a high anti-interference capability and low background due to the presence of both GO and black hole quencher (BHQ) as a dual-quencher that reduces the background in live cell imaging due to resonance energy transfer (RET). We then employed the DNA-GO nanocomplex probe for simultaneous detection of miR-630 and miR-21 and also for simultaneous in situ dynamic monitoring of intracellular miR-630 and miR-21 in apoptotic cells. We discovered that miR-630 expression was up-regulated during the first 120 min. This simple but powerful protocol has great potential in precise detection and imaging of various substances in complex biological samples with improved accuracy.

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confidence: 99%

Dual Energy Transfer-Based DNA/Graphene Oxide Nanocomplex Probe for Highly Robust and Accurate Monitoring of Apoptosis-Related microRNAs

et al. 2020

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“…Participation of complement in acute-or-chronic liver failure, although a very likely event, remains to be explored. In situ imaging with DNA hybridization and fluorescence could focus on complement in liver disease [ 67 ].…”

Section: Complement Activation In Alcoholic Liver Diseases (Ald)mentioning

confidence: 99%

The complement system in liver diseases: Evidence-based approach and therapeutic options

Lung

Sakem

Risch

et al. 2019

Journal of Translational Autoimmunity

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Complement is usually seen to largely originate from the liver to accomplish its tasks systemically – its return to the production site has long been underestimated. Recent progress in genomics, therapeutic effects on complement, standardised possibilities in medical laboratory tests and involvement of complosome brings the complement system with its three major functions of opsonization, cytolysis and phagocytosis back to liver biology and pathology. The LOINC™ system features 20 entries for the C3 component of complement to anticipate the application of artificial intelligence data banks algorythms of which are fed with patient-specific data connected to standard lab assays for liver function. These advancements now lead to increased vigilance by clinicians. This reassessment article will further elucidate the distribution of synthesis sites to the three germ layer-derived cell systems and the role complement now known to play in embryogenesis, senescence, allotransplantation and autoimmune disease. This establishes the liver as part of the gastro-intestinal system in connection with nosological entities never thought of, such as the microbiota-liver-brain axis. In neurological disease etiology infectious and autoimmune hepatitis play an important role in the context of causative viz reactive complement activation. The mosaic of autoimmunity, i.e. multiple combinations of the many factors producing varying clinical pictures, leads to the manifold facets of liver autoimmunity.

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“…However, the improved detection sensitivity of isothermal amplification methods may be compromised by the requirement of DNA polymerase or other enzymes and by poor reproducibility. Therefore, enzyme-free amplification techniques are more attractive for biological and biomedical applications. − …”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive Detection of miR-21 through Target-Activated Catalytic Hairpin Assembly of X-Shaped DNA Nanostructures

Chai

et al. 2021

Anal. Chem.

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MicroRNAs (miRNAs) are found in extremely low concentrations in cells, so highly sensitive quantitation is a great challenge. Herein, a simple dual-amplification strategy involving target-activated catalytic hairpin assembly (CHA) coupled with multiple fluorophores concentrated on one X-shaped DNA is reported. In this strategy, four hairpin probes (H1, H2, H3, and H4) are modified with FAM and BHQ1 at both sticky ends, while a circulating hairpin probe (H0) is used to activate CHA circuits once it binds to complementary sequences in the target miR-21 (T). The powerful dual-amplification cascades in Förster resonance energy transfer (FRET)-based nonenzymatic nucleic acid circuits are triggered by T–H0-activated formation of the X-shaped DNA nanostructure, freeing T–H0 for the next CHA reaction cycle. CHA circuits increase the fluorescence due to the wide distance between FAM and BHQ1 in the formed X-shaped DNA nanostructure, resulting in signal amplification and highly sensitive detection of miR-21, with a limit of detection (LOD, 3σ) of 0.025 nM, which is 25.6 or 57.6 times lower than that obtained through a single-amplification strategy without multiple fluorophores on one X-shaped DNA or CHA circuit. Furthermore, this cascade reaction was completed in 45 min, effectively avoiding target degradation. This new enzyme-free signal amplification strategy holds promising potential for sensitively detecting different DNA or RNA sequences by simply adapting the fragment of the H0 sequence complementary to the target.

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Dual Energy Transfer-Based Fluorescent Nanoprobe for Imaging miR-21 in Nonalcoholic Fatty Liver Cells with Low Background

Cited by 32 publications

References 41 publications

Dual Energy Transfer-Based DNA/Graphene Oxide Nanocomplex Probe for Highly Robust and Accurate Monitoring of Apoptosis-Related microRNAs

Dual Energy Transfer-Based DNA/Graphene Oxide Nanocomplex Probe for Highly Robust and Accurate Monitoring of Apoptosis-Related microRNAs

The complement system in liver diseases: Evidence-based approach and therapeutic options

Highly Sensitive Detection of miR-21 through Target-Activated Catalytic Hairpin Assembly of X-Shaped DNA Nanostructures

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