DNA logic nanomachine for the accurate identification of multiple microRNAs in tumor cells
The use of dynamic DNA logic circuits for disease diagnosis at the molecular level plays a considerable role in biomedical fields. Nevertheless, how to create programmable nanomachines based on molecular logical gates to accurately identify multiple biomarkers from tumor cells remains a pivotal challenge. Herein, we developed a DNA-based nanomachine for analyzing and imaging multiple microRNAs (miRNAs) in cancerous cells with a logical AND operation. The triangular prism design of DNA nanomachine improved its performance in living cell research with high stability and served as a modularized framework for toehold-mediated strand displacement reactions and catalytic hairpin assembly circuits.
The results suggested that the nanomachine could efficiently enter cells with great biocompatibility and rapidly recognize the correct biomolecules with high sensitivity. The well-designed DNA-logic gate nanomachine enabled accurate diagnosis on multiple miRNA patterns in different cell lines and differentiation of aberrant expression in target cells, which provided a novel possibility for intelligent disease diagnosis using smart nanomachines at the molecular level.
Chemicals and materials
The sequences of synthesized oligonucleotides are listed in Table S1, the reagents and materials in the experiments are described in the Supplementary Material.
DNA prism probe was prepared through adding equimolar nucleic acid strands of S1, S2, S3, H1-T, H2-T/H2-T-FQ, Capture-T, Trigger, Apt-1, Apt-2 and Apt-3 in 1 × TAE-Mg2+ buffer (40 mM Tris buffer, 1 mM EDTA, 12.5 mM MgAc2, pH 8.0) to the final concentration of 1 μM. The mixture was then subjected to a thermal annealing process (Table S2)
The working principle of self-assembled DNA nanomachines for simultaneous identification and imaging of multi-miRNAs is exhibited in Scheme 1. Initially, the 3D DNA nanomachine was one-pot self-assembled from ten nucleic acid strands, of which three long strands (S1, S2, S3) for assembling the triangular prism framework, two strands (Capture-T and Trigger) for synthesizing an “Anchor” domain to identify target miRNAs, two hairpin strands (H1-T and H2-T) as CHA substrate strands, and three.
In this work, DNA-logic gate nanomachines activated by multi-target miRNAs within cells were successfully designed using a DNA triangular prism as the nano-vehicle and the CHA cascade circuit as a signal amplifier. After internalization, DNA nanomachines made logical responses and the Trigger strand could be efficiently released via TMSD in the presence of miRNA-21, miRNA-155, and miRNA-10b simultaneously. Whereafter, CHA was activated by dissociative Trigger strand to generate amplified.
tumor microenvironment, cancer stem cells, metastasis, angiogenesis, immune evasion, oncogenes, tumor suppressor genes, apoptosis resistance, epithelial-mesenchymal transition, tumor heterogeneity, cell proliferation, hypoxia, tumor markers, neoplastic transformation, chemotherapy resistance, cell signaling pathways, tumorigenesis, genomic instability, cancer immunotherapy, tumor metabolism
#TumorCells, #CancerResearch, #Oncology, #Metastasis, #TumorMicroenvironment, #CancerStemCells, #TumorImmunology, #CancerTherapy, #ApoptosisResistance, #TumorMarkers, #Angiogenesis, #CellProliferation, #CancerGenetics, #EMT, #TumorHeterogeneity, #TumorMetabolism, #Chemoresistance, #PrecisionOncology, #CancerSignaling, #GenomicInstability
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