通讯机构:
[Lin Yuan] S;State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, P. R. China
摘要:
Herein, a novel two-photon ratiometric fluorescence assay was proposed for monitoring endogenous steroid sulfatase (STS) activity, which could be applied for the ratiometric imaging of STS activity in the endoplasmic reticulum of living cells and tissues and also could be used to distinguish estrogen-dependent tumor cells from other types of cells.
摘要:
The Au−S bond is the classic way to functionalize gold nanoparticles (AuNPs). However, cleavage of the bond by biothiols and other chemicals is a long-standing problem hindering practical applications, especially in cells. Instead of replacing the thiol by a carbene or selenol for stronger adsorption, it is now shown that the Pt−S bond is much more stable, fully avoiding cleavage by biothiols. AuNPs were deposited with a thin layer of platinum, and an AuNP@Pt-S nanoflare was constructed to detect the miRNA-21 microRNA in living cells. This design retained the optical and cellular uptake properties of DNA-functionalized AuNPs, while showing high-fidelity signaling. It discriminated target cancer cells even in a mixed-cell culture system, where the Au-S based nanoflare was less sensitive. Compared to previous methods of changing the ligand chemistry, coating a Pt shell is more accessible, and previously developed methods for AuNPs can be directly adapted.
通讯机构:
[Qing, ZH; Yang, RH] C;Changsha Univ Sci & Technol, Hunan Prov Key Lab Mat Protect Elect Power & Tran, Hunan Prov Engn Res Ctr Food Proc Aquat Biot Reso, Sch Chem & Food Engn, Changsha 410114, Peoples R China.
摘要:
Enzyme-free amplification techniques based on dynamic DNA self-assembly (DDSA) have recently been developed for the in situ detection of mRNA in living cells. However, signal generation in traditional DDSA amplifiers is mainly dependent on the random diffusion of dissociative probes in a bulk solution, which is generally accompanied by poor kinetics and interference from complex biological systems. In this work, a new amplifier based on the design of an intramolecular catalytic hairpin assembly (intra-CHA) is proposed for the FRET imaging of mRNA in living cells. Compared with that in the free catalytic hairpin assembly (free-CHA), probes H1 and H2 in intra-CHA were simultaneously fixed on a DNA tetrahedron. The distance between them was closer, the local concentration of H1 and H2 in intra-CHA was theoretically approximately 808-times higher than that in free-CHA, and the initial reaction rate was enhanced 15.6 fold. Due to the spatial confinement effect, the reaction kinetics for target-catalyzed signal generation were significantly improved. By virtue of the three-dimensional nanostructure, H1 and H2 in the intra-CHA amplifier entered cells without any transfection or nanocarrier, and the probes and their products were free from biological interference, providing much higher signal stability for the reliable imaging of mRNA in living cells.
摘要:
For the first time it isdemonstrated that sulfhydryl compounds can suppress longitudinal etching of gold nanorods via consuming oxidizers, which provides a new signaling mechanism for colorimetric sensing. As a proof of concept, a colorimetric assay is developed for detecting organophosphorus pesticides, which are most widely used in modern agriculture to improve food production but with high toxicity to animals and the ecological environment. Triazophos was selected as a model organophosphorus pesticide. In the absence of triazophos, the active acetylcholinesterase can catalyze the conversion of acetylthiocholine iodide to thiocholine whose thiol group can suppress the I(2)-induced etching of gold nanorods. When triazophos is present, the activity of AchE is inhibited, and I(2)-induced etching of gold nanorods results in triazophos concentration-dependent color change from brown to blue, pink, and red. The aspect ratio of gold nanorods reduced with gradually blue-shifted longitudinal absorption. There was a linear detection range from 0 to 117nM (R(2) = 0.9908), the detection limit was 4.69nM, and a good application potential was demonstrated by theassay of real water samples. This method will not only contribute to public monitoring of organophosphorus pesticides but also has verified a new signaling mechanism which will open up a new path to develop colorimetric detection methods. It has been first found that sulfhydryl compounds can suppress longitudinal etching of gold nanorods (AuNRs) via consuming oxidizers, which provides a new signaling mechanism for colorimetric sensing. As a proof of concept, a colorimetric assay is developed for sensitively detecting organophosphorus pesticides (OPs). It will not only contribute to public monitoring of OPs but also has verified a new signaling mechanism which will open up a new path to develop multicolor colorimetric methods.
摘要:
As VEGF mRNA is an endothelial cell-specific mitogen and a key regulator of angiogenesis in a variety of physiological and pathological processes, high expression levels of VEGF messenger RNA (mRNA) contribute to VEGF-driven angiogenesis in the hypoxic areas of solid tumors and then disrupt the vascular barrier, which may potentiate tumor cell extravasation. Thus, monitoring the changes in VEGF mRNA is necessary to understand the genetic programme under hypoxic conditions and thus facilitate risk assessment and risk reduction in hypoxic environments. Herein, a new fluorescent nanoprobe based on azoreductase-responsive functional metal-organic frameworks (AMOFs) was developed to realize the imaging of VEGF mRNA under hypoxic conditions. Since the azobenzene units in the AMOFs can be reduced to amines by the highly expressed azoreductase in an oxygen-deficient environment, the VEGF mRNA-targeted molecular beacon (MB), which is adsorbed on the surface of AMOFs via electrostatic interactions, can be released due to the structural damage of AMOFs. Moreover, TAMRA (carboxytetramethyl-rhodamine, donor) and Cy5 (acceptor) were close to each other due to the stem-loop conformation of MB, thus inducing high fluorescence energy resonance transfer (FRET) efficiency. Upon the addition of VEGF mRNA, the hybridization of VEGF mRNA destroyed the stem-loop conformation of MB, and then, the two fluorophores labeled on MB were separated with low FRET efficiency. This constructed fluorescent nanoprobe enables the quantitative analysis and in situ imaging of the VEGF mRNA level in living cells under hypoxic conditions. We expect that it will offer a potentially rich opportunity to understand the physiological processes of genetic programme.
摘要:
Fluorescence molecular imaging has attracted increasing attention due to its various advantages. Lots of fluorophores have been developed to meet various molecular imaging needs. However, it is still inconvenient due to the lack of excellent fluorophores with an optically tunable group for biological molecular imaging. Here a new platform of a versatile long wavelength fluorophore with an optically tunable hydroxyl group was successfully developed by regulating molecular planarity and the twisted intramolecular charge transfer effect with a protected and deprotected hydroxyl group approach via "step by step" modifying strategy. As an excellent representative of this new type of fluorophore, LDOH-4 possesses good chemical and optical properties and shows a potential application prospect. As a proof-of-concept, a nitroreductase-activated TP fluorescent probe LDO-NTR was designed, which not only sensitively recognizes NTR with more than 310-fold response signal enhancement in vitro but also tracks NTR in a hypoxia tumor mouse model in vivo by using two-photon imaging. It is hopeful that the long wavelength fluorophore with the optically tunable hydroxyl group can serve as a useful platform to extend capable detection tools in biological chemistry and biomedical applications.
作者机构:
[Zheng, Jing; Zhu, Cong; Chen, Shiya] Hunan Univ, Coll Chem & Chem Engn, State Key Lab Chemo Biosensing & Chemometr, Changsha 410082, Hunan, Peoples R China.;[Huang, Fujian] China Univ Geosci, Fac Mat Sci & Chem, Minist Educ, Engn Res Ctr Nanogeomat, Wuhan 430074, Hubei, Peoples R China.;[Yang, Ronghua] Changsha Univ Sci & Technol, Sch Chem & Biol Engn, Changsha 410076, Hunan, Peoples R China.;[Yang, Jinfeng] Cent S Univ, Hunan Canc Hosp, Affiliated Canc Hosp, Xiangya Sch Med, Changsha 410083, Hunan, Peoples R China.
通讯机构:
[Zheng, Jing] H;[Huang, Fujian] C;Hunan Univ, Coll Chem & Chem Engn, State Key Lab Chemo Biosensing & Chemometr, Changsha 410082, Hunan, Peoples R China.;China Univ Geosci, Fac Mat Sci & Chem, Minist Educ, Engn Res Ctr Nanogeomat, Wuhan 430074, Hubei, Peoples R China.
摘要:
A new triplex-functionalized DNA tetrahedral nanoprobe is proposed herein for monitoring pH and messenger RNA (mRNA) in living cells. Different from traditional DNA tetrahedron-based nanoprobes, DNA triplex was employed to serve as important conformational conversion elements. Inspired by the low extracellular pH in tumor cells, the mRNA-targeted H1 and H2 were stably assembled on the extended short hairpin probes of DNA tetrahedron via Hoogsteen bonding to form DNA triplex. Due to the high intracellular pH and presence of target mRNA, hybridization chain reaction (HCR) was triggered between H1 and H2 which were released from the dissociation of DNA triplex, and the generated long double-stranded DNA activated a Foster resonance energy transfer (FRET) signal indicating target mRNA expression even at very low contents. By combining the distinguishing feature of DNA triplex structure (pH-responsive) and HCR (signal amplification), sensitive imaging of intracellular pH and tumor-related mRNA can be realized. As a further application, dynamic imaging of intracellular pH and mRNA during "mitochondria-dependent" pathway apoptosis was successfully achieved in human breast cancer cells, which indicated huge potential of our proposed nanoprobe in early diagnosis and treatment of diseases.
摘要:
A colorimetric and visual assay is described for the herbicide aminotriazole (ATZ). It is based on the etching of gold nanorods (AuNRs) by iodine which is formed on oxidation of iodide via H2O2. Longitudinal etching of the AuNRs occurs quickly and is accompanied by a color change from dark blue to red. In the absence of ATZ and the presence of active catalase (CAT), H2O2 is quickly decomposed into water, and the AuNRs will not be etched. In the presence of ATZ, CAT is partially deactivated, and this affects the amount of available H2O2 and, consequently, of the iodine. Hence, the color is significantly changed. The color changes can be easily detected with bare eyes. The assay has a linear response in the 5 to 70 μM concentration range, with a detection limit of 1.3 μM and high selectivity for ATZ. It was applied to the determination of ATZ in water and food samples. Graphical abstractA multicolor colorimetric method is developed for aminotriazole (ATZ) detection based on catalase (CAT) deactivation-dependent longitudinal etching of gold nanorods (AuNRs). The color signals can be visually identified. Good detection performances and capability for evaluating ATZ level in water and food samples is demonstrated.
摘要:
We report here a two-photon nanoprobe for the detection of RNase H activity in living cells and ex vivo tissues by combining a two-photon dye with a spherical nucleic acid (SNA) featuring a DNA/RNA duplex corona and a gold nanoparticle core.