Intracellular activated logic nanomachines based on framework nucleic acids for low background detection of microRNAs in living cells

Abstract

DNA molecular machines based on DNA logic circuits show unparalleled potential in precision medicine. However, delivering DNA nanomachines into real biological systems and ensuring that they perform functions specifically, quickly...

Logic Signal Amplification System for Sensitive Electrochemiluminescence Detection and Subtype Identification of Cancer Cells

“…MCF-10A was selected as the control sample, with classical qRT-PCR technology employed for verification (Figure S11). The trend of miR-21 expression levels in MCF-10A, MCF-7, and HeLa cells matched that detected by our constructed biosensor (Figure A), aligning with the literature previously reported . The aforementioned results displayed that the constructed biosensor was reliable in the detection of cell lysates and could be utilized to differentiate between cancer and normal cells.…”

“…Molecular logic gates, comparable to silicon-based electronic gates in integrated circuits, facilitate a range of computations at the molecular scale. , DNA logic gates are capable of responding to single or multiple targets, yielding outputs after logic processing. , The AND gates in DNA computation aid in mitigating the effects of phenotypic heterogeneity, compared to single-input detection. , By employing DNA logic input–output signal patterns, it is possible to conduct a multiparametric analysis of microRNAs (miRNAs), thereby enabling the precise identification of various cancer cell subtypes. , However, due to the high sequence homology and low abundance of nucleic acid, there are great challenges in the sensitive detection of cancer-related nucleic acids. , Benefiting from the inherent advantages of being enzyme-free, isothermal and programmable, isothermal amplification techniques have received much attention. − Catalytic hairpin assembly (CHA) represents a method for targeted catalytic amplification, wherein an initiator may activate two or more hairpins to execute a sequence of cascading assembly reactions, yielding distinct DNA nanostructures. − However, the traditional nonenzymatic DNA reactants have low collision efficiency in the free diffusion state of solution, resulting in slow kinetics and relatively weak detection ability. − Therefore, developing robust and powerful systems to improve the problem is of significant importance for achieving a weak-input-strong-output model in DNA logic gates.…”

Remotely Activated DNA Probe System for the Detection and Imaging of Dual miRNAs

“…To overcome these shortcomings, considerable DNA-based miRNA detection methods have been developed. − To enhance performance, the construction of stimulus-responsive DNA nanoplatforms is crucial for the accurate detection and imaging of miRNAs. Triggers such as enzymes, pH, and metal ions are developed to have more precise control of the system. − Light is frequently utilized as an effective tool for various regulations because it can be simply and accurately regulated, allowing for exact modulation of biosensor activity. − Once it reaches the target location, it can be selectively activated to monitor miRNA with great precision.…”

“…Related research has grown tremendously during the past few years . It has been employed for miRNA detection, ,, subcellular organelle targeting, and imaging using intracellular cues ,, such as H + , endonuclease, membrane proteins, logic-gated sensor designs, tumor therapy, and so on. The concept was further extended to other combinations of materials, such as SiO 2 , AuNPs, or MOFs for biomarker detection.…”

Endogenous AND Logic DNA Nanomachine for Highly Specific Cancer Cell Imaging

“…Specific imaging of cancer cell based on biomarkers is of high significance to achieve the early cancer diagnosis and the reduction of disease related mortality. − The combination of precise base-pairing, excellent biocompatibility, and the high programmability has rendered DNA circuits excellent devices for biomedical imaging in living cells. − Particularly, DNA circuits that could ingeniously integrate programmable DNA self-assembly and functional modules of signal amplification have been constructed for low-abundance biomarkers imaging. − As a typical isothermal DNA circuit, toehold-mediated strand displacement (TMSD) reaction has been developed for amplification detection of biomarkers of interest. , Besides, spatial-confinement effect limited the substrates into a nanoscale for high local concentration, which significantly enhanced the operating efficiency of those DNA circuits. ,− Based on the principle, some emerging DNA circuits, including localized TMSD circuit, were developed. , Despite the improvements in the sensitivity of these methods, cancer cell imaging specificity is still constrained, mainly owing to the signal leakages from the confined space , and the false positive associated with single-biomarker detection, because most biomarkers overexpressed in cancer cells also exist in normal cells, such as miR-21. , Accordingly, establishing multiple activation DNA circuits with specific control to achieve highly reliable cancer cell imaging is needed.…”