NEW JOURNAL OF CHEMISTRY
·
2018
·Vol. 42
·期 17
·14325-14331
·
IF 3.201
Ruiqing Feng,Longlong Li,Bing Li,Jinhui Li,Dan Peng,Yitao Yu,Qiuhong Mu,Ning Zhao,Xiaoqiang Yu,Zhenhua Wang
We have synthesized a two-photon organic small-molecule probe (DMI), which has a considerable two-photon absorption cross section (δ) value and selectively stained endogenous RNA in the nucleolus and the cytoplasm of living cells with outstanding membrane permeability. The titrations, fluorescence imaging and RNase digest test indicate markedly higher affinity for endogenous RNA. The double assay of DMI and SYTOX Blue nucleic acid stain (S-11348) showed that the probe possesses excellent membrane permeability to living cells, and a co-staining experiment with the classic DNA-staining dye DAPI indicated that it has excellent counterstain compatibility. Moreover, DMI exhibited good photostability when continuously illuminated by constant femtosecond pulses. These results demonstrate that DMI is an attractive two-photon fluorescent probe for visualizing RNA in living cells.
研究领域:
生物化学
·
细胞生物学
·
分子影像学
✓ 已验证
产品关联
DOI
高影响因子
MOLECULES
·
2023
·Vol. 28
·期 5
·2149
·
IF 4.6
Zhe Chen,Zhuoyi Wang,Yihua Yuan,Bo Liu,Jiangbo Yu,Zhiwen Wei,Keming Yun
DNA self-assembled fluorescent nanoprobes have been developed for bio-imaging owing to their high resistance to enzyme degradation and great cellular uptake capacity. In this work, we designed a new Y-shaped DNA fluorescent nanoprobe (YFNP) with aggregation-induced emission (AIE) characteristic for microRNA imaging in living cells. With the modification of the AIE dye, the constructed YFNP had a relatively low background fluorescence. However, the YFNP could emit a strong fluorescence due to the generation of microRNA-triggered AIE effect in the presence of target microRNA. Based on the proposed target-triggered emission enhancement strategy, microRNA-21 was detected sensitively and specifically with a detection limit of 122.8 pM. The designed YFNP showed higher bio-stability and cell uptake than the single-stranded DNA fluorescent probe, which has been successfully applied for microRNA imaging in living cells. More importantly, the microRNA-triggered dendrimer structure could be formed after the recognition of target microRNA, achieving a reliable microRNA imaging with a high spatiotemporal resolution. We expect that the proposed YFNP will become a promising candidate for bio-sensing and bio-imaging.Keywords:DNA fluorescent nanoprobes;Y-shaped;aggregation-induced emission;microRNA;live-cell imaging
研究领域:
生物化学
·
分子生物学
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Yanfei Zhang,Jun Chen,Huihui Yang,Wen Yin,Chunrong Li,Yuzhi Xu,Si-Yang Liu,Zong Dai,Xiaoyong Zou
Sensitive imaging of intracellular microRNA (miRNA) in living cells is of great significance. Isothermal hybridization chain reaction (HCR)-based methods, although have been widely used to monitor intracellular low-abundance miRNA, are still subjected to the challenges of limited signal amplification efficiency and compromised imaging resolution. In this work, we design a light-controlled recruitable HCR (LCR-HCR) strategy that enables us to well overcome these limitations. Exosomes as delivery and recruitment vehicles are modified with three cholesterol-modified hairpins (H1, H2, and H3), in which H1 is for anchoring target miRNA and H2 and H3 with photocleavable linkers (PC-linkers) are designed for spatiotemporal HCR. By controllably releasing probes with high local concentrations to efficiently trigger HCR and further recruiting the generated double-stranded DNA (dsDNA) polymers instead of dispersion in the cytoplasm, the LCR-HCR method can significantly improve the imaging contrast by confining all of the reactants on exosome vehicles. For a proof-of-concept demonstration, the miR-21 was analyzed by LCR-HCR with a limit of detection (LOD) down to 3.3 pM (corresponding to 165 amol per 50 μL) in vitro and four times higher response than traditional HCR in vivo. In general, the LCR-HCR method provides a powerful tool for sensitive miRNA imaging in living cells and cancer diagnosis.
研究领域:
生物技术
·
分子生物学
·
癌症研究
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细胞生物学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Zizhong Yang,Birong Liu,Ting Huang,Bao-Ping Xie,Wen-Jun Duan,Min-Min Li,Jin-Xiang Chen,Jun Chen,Zong Dai
Sensitive and specific imaging of microRNA (miRNA) in living cells is of great value for disease diagnosis and monitoring. Hybridization chain reaction (HCR) and DNAzyme-based methods have been considered as powerful tools for miRNA detection, with low efficient intracellular delivery and limited amplification efficiency. Herein, we propose a Hairpins@MnO2 nanosystem for intracellular enzyme-free exponential amplification for miRNA imaging. The enzyme-free exponential amplification is based on the synergistic cross-activation between HCR and DNAzymes. The MnO2 nanosheets were employed as the carrier of three kinds of hairpin DNA probes and further provided appropriate Mn2+ as DNAzyme cofactors in the living cell. Upon entering cells and in the presence of highly expressed glutathione (GSH) in tumors, MnO2 is reduced to release Mn2+ and the three kinds of hairpin DNA probes. In the presence of target miRNA, the released hairpin DNA H1 and H2 probes self-assemble via HCR into the wire-shaped active Mn2+-based DNAzymes which further catalyze the cleavage of H3 to generate numerous new triggers to reversely stimulate HCR amplifiers, thus offering tremendously amplified Förster resonance energy transfer readout. The method has a detection limit of 33 fM, which is 2.4 × 104 times lower than that of the traditional HCR system. The developed method also has a high specificity; even miRNAs with a single base difference can be distinguished. Live cell imaging experiments confirmed that this Hairpins@MnO2 nanosystem allows accurate differentiation of miRNA expression of cancer cells and normal cells. The method holds great potential in biological research of nucleic acids.
研究领域:
生物化学
·
分子生物学
·
纳米技术
·
癌症诊断
✓ 已验证
DOI
PubMed
高影响因子
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