Dataset Information

Fluorescence-coded DNA Nanostructure Probe System to Enable Discrimination of Tumor Heterogeneity via a Screening of Dual Intracellular microRNA Signatures in situ.

ABSTRACT: Since the delivery kinetics of different cell types are different, the signal from the target cell is greatly affected by the noise signal of the diagnostic system. This is a major obstacle hindering the practical application of intracellular diagnostic systems, such as tumor heterogeneity. To address these issues, here we present a microRNA detection platform using fluorescence-encoded nanostructured DNA-based probes. The nanostructured DNA was designed to include molecular beacons for detecting cytosolic microRNA as well as additional fluorophores. When the intracellular diagnostic system is delivered, fluorescence signals are generated by the molecular beacons, depending on the concentration of the target microRNA. The fluorescence signals are then normalized to the intensity of the additional fluorophore. Through this simple calculation, the concentration of intracellular microRNA can be determined without interference from the diagnosis system itself. And also it enabled discrimination of microRNA expression heterogeneity in five different breast cancer cell lines.

SUBMITTER: Shin SW

PROVIDER: S-EPMC5647416 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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Fluorescence-coded DNA Nanostructure Probe System to Enable Discrimination of Tumor Heterogeneity via a Screening of Dual Intracellular microRNA Signatures in situ.

Shin Seung Won SW Lee Byoung Sang BS Yang Kisuk K Amornkitbamrung Lunjakorn L Jang Min Su MS Ku Bo Mi BM Cho Seung-Woo SW Lee Jung Heon JH Bae Hojae H Oh Byung-Keun BK Ahn Myung-Ju MJ Lim Yong Taik YT Um Soong Ho SH

Scientific reports 20171018 1

Since the delivery kinetics of different cell types are different, the signal from the target cell is greatly affected by the noise signal of the diagnostic system. This is a major obstacle hindering the practical application of intracellular diagnostic systems, such as tumor heterogeneity. To address these issues, here we present a microRNA detection platform using fluorescence-encoded nanostructured DNA-based probes. The nanostructured DNA was designed to include molecular beacons for detectin ...[more]

PMID: 29044199

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Project description:Prorocentrum donghaiense is a common but dominant harmful algal bloom (HAB) species, which is widely distributed along the China Sea coast. Development of methods for rapid and precise identification and quantification is prerequisite for early-stage warning and monitoring of blooms due to P. donghaiense. In this study, sequences representing the partial large subunit rDNA (D1-D2), small subunit rDNA and internal transcribed spacer region (ITS-1, 5.8S rDNA and ITS-2) of P. donghaiense were firstly obtained, and then seven candidate DNA probes were designed for performing fluorescence in situ hybridization (FISH) tests on P. donghaiense. Based on the fluorescent intensity of P. donghaiense cells labeled by the DNA probes, the probe DP0443A displayed the best hybridization performance. Therefore, a PNA probe (PP0443A) analogous to DP0443A was used in the further study. The cells labeled with the PNA probe displayed more intensive green fluorescence than that labeled with its DNA analog. The PNA probe was used to hybridize with thirteen microalgae belonging to five families, i.e., Dinophyceae, Prymnesiophyceae, Raphidophyceae, Chlorophyceae and Bacillariophyceae, and showed no visible cross-reaction. Finally, FISH with the probes PP0443A and DP0443A and light microscopy (LM) analysis aiming at enumerating P. donghaiense cells were performed on the field samples. Statistical comparisons of the cell densities (cells/L) of P. donghaiense in the natural samples determined by FISH and LM were performed using one-way ANOVA and Duncan's multiple comparisons of the means. The P. donghaiense cell densities determined by LM and the PNA probe are remarkably higher than (p<0.05) that determined by the DNA probe, while no significant difference is observed between LM and the PNA probe. All results suggest that the PNA probe is more sensitive that its DNA analog, and therefore is promising for the monitoring of harmful algal blooms of P. donghaiense in the future.

Dataset Information

Fluorescence-coded DNA Nanostructure Probe System to Enable Discrimination of Tumor Heterogeneity via a Screening of Dual Intracellular microRNA Signatures in situ.

Publications

Fluorescence-coded DNA Nanostructure Probe System to Enable Discrimination of Tumor Heterogeneity via a Screening of Dual Intracellular microRNA Signatures in situ.

Similar Datasets

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