Functional nucleic acids (FNAs) enhance loop-mediated isothermal amplification (LAMP) for next-generation diagnostics. This review structures FNAs across the LAMP workflow: target enrichment and specificity locking, isothermal amplification with real-time monitoring, logic gating, and catalytic/electrochemical signal amplification. These strategies are driven by molecular recognition, catalytic signal generation, dynamic switching, structural stabilization, and chemical modification. Specifically, aptamer-mediated magnetic capture and PNA/LNA clamping enable target enrichment and suppress nonspecific amplification. During isothermal amplification, molecular beacons leverage strand displacement for real-time fluorescence generation. For logic gating, transcription factor-based AND gates and riboswitches achieve conditional signal transduction. For catalytic and electrochemical amplification, DNAzymes, i-motifs, and G-quadruplexes provide catalytic conversion, pH-responsive colorimetry, and electrochemical readouts. Integration with functional materials further boosts sensitivity and point-of-care compatibility. Demonstrated applications include detecting foodborne pathogens, toxins, allergens, and GMOs for rapid on-site screening. Future directions focus on intelligentization, automation, and portability for resource-limited and industrial settings.

Fluorescent RNA aptamers offer promising opportunities for next-generation biosensing but are often limited by low signal-to-background ratios and unstable folding kinetics. In this work, a label-free Forster resonance energy transfer (FRET)-enhanced fluorescent artificial RNA condensate (F-FARCON) is developed for small-molecule sensing, leveraging neutral molecular crowders (e.g., polyethylene glycol 8K), and RNA structural motifs to induce multivalent interactions and drive dynamic self-assembly. As a demonstration, a label-free FRET system is constructed by integrating a histamine-responsive RNA aptamer with thioflavin T (ThT) as the fluorescence donor, which increases the signal-to-noise ratio while reducing sequence complexity and production costs. Molecular crowders optimize the thermodynamic environment of RNA-ligand and RNA-RNA multivalent interactions, thereby improving folding stability, signal amplitude (dynamic range of up to similar to 970-fold), and target affinity. The platform exhibits fast kinetics (<15 min), an adjustable detection range (0.1-200 and 5-1000 & micro;M), and high sensitivity (limit of detection, 15.36 nM), with robust performance in complex biological matrices. The platform is further integrated into a freeze-dried paper-based portable device that enables dual-channel fluorescence readout for on-site rapid detection without sophisticated instrumentation. To further validate the modularity of F-FARCON beyond histamine, we reprogrammed the recognition module to target S-adenosyl-L-methionine (SAM), achieving nanomolar limits of detection. By linking crowding-guided assembly to hierarchical photophysical enhancement and analytical performance, the work delineates a generalizable aggregate-science route to versatile, low-cost, and field-deployable fluorescence sensing across food safety, environmental monitoring, and biomedical diagnostics.

Cronobacter sakazakii (C. sakazakii), an opportunistic pathogenic bacterium resistant to adverse environments, can induce meningitis and sepsis in infants with high mortality, which necessitates ultra-sensitive detection. In this study, the core binding region was derived by tailoring existing C. sakazakii aptamer, and novel aptamer screening libraries were designed based on this core sequences with high affinity and specificity. Additionally, we performed the first screening of C. sakazakii DNAzyme and explored its binding sites with the aid of molecular docking. Based on the aptamer, DNAzyme, G-quadruplex, and RCA reaction, an ultra-sensitive visual C. sakazakii sensor was developed. This sensor achieves C. sakazakii enrichment via aptamer-magnetic bead conjunction and converts bacterial count to nucleic acid count signals through DNAzyme. Rolling circle amplification (RCA) generates massive G-quadruplexes for exponential signal amplification; the complex formed by G-quadruplex and hemin catalyzes TMB chromogenesis to complete the final visual signal output. Under optimized conditions, the sensor exhibits a linear relationship in the range of 9 × 100 ∼ 9 × 103 CFU/mL, with a linear equation of y = 0.2282LgC + 0.1957 (R2=0.9992) and a limit of detection (LOD) of 2 CFU/mL. In summary, this study developed an ultra-sensitive sensor based on multiple functional nucleic acids for C. sakazakii detection.

Vitamin E can exert either a cancer preventive effect or improve the therapeutic efficacy of chemotherapeutic drugs against multiple types of cancer. Ample evidence suggests that the cancer preventive activity of vitamin E is form-dependent; however, it is not clear whether its chemosensitization effect is also influenced by its forms. The objectives of this study were to investigate whether the eight natural forms of vitamin E produced differential sensitization effects on cancer chemotherapeutic drugs and to address whether the chemosensitization effect of vitamin E was associated with its inhibitory effect on programmed cell death ligand 1 (PD-L1) signaling. We carried out a comparative evaluation of the chemosensitization effect of eight vitamin E forms using paclitaxel as a representative therapeutic drug and breast/prostate cancer as the representative types of cancer. Results showed that the sensitization effect of vitamin E on chemotherapeutic drugs was also form-dependent, with δ-tocotrienol (δ-T3) as the most effective one for sensitizing breast and prostate cancer cells to paclitaxel, mechanistically associated with its ability to suppress PD-L1-mediated tumor-promoting signaling. The findings provided novel insights into understanding the sensitization effect of vitamin E and its related mechanisms and support that δ-T3 is the best candidate as an enhancer of taxanes among the eight forms.

β-Lactoglobulin (BLG), the principal allergen in bovine milk, exhibits strong immunogenicity and structural stability, posing notable challenges to food safety. Accurate detection of BLG is critical for allergen labeling and consumer protection. This review systematically summarizes molecular recognition strategies for BLG detection. It defines eight recognition modes and three binding sites, and categorizes seven recognition elements, including antibodies, aptamers, specific binding peptides, molecularly imprinted polymers, fluorogenic probes, metal nanozymes, and primers, based on binding mechanisms. Additionally, direct strategies utilizing BLG's intrinsic signatures, such as mass spectrometry and spectroscopy, are reviewed. Detection platforms are evaluated in terms of sensitivity, specificity, and applicability. Future perspectives highlight advances in intelligent sensing technologies, including integration with nanomaterials, microfluidics, and digital analysis. Emphasis is placed on achieving high performance under complex food conditions. This review provides a concise reference for developing next-generation BLG biosensors with improved accuracy, portability, and suitability for real-world allergen monitoring.

Over the past decade, the study of DNA-templated metal nanoclusters (DNA-MNCs) has made rapid progress. It demonstrates broad application prospects in the fields of biosensing, medical diagnostics, and environmental monitoring. Despite the significant achievements of DNA-MNCs in optical properties, little study has been conducted on the catalytic properties of DNA-MNCs. We have successfully synthesized DNA-Au/Pt bimetallic NCs through a chemical reduction method, which exhibit excellent peroxidase-like (POD-like) activity. Due to the synergistic effect between the bimetals and an efficient electron transfer mechanism, their catalytic activity far surpasses that of monometallic NCs, and their affinity for hydrogen peroxide (H2O2) is even superior to horseradish peroxidase (HRP). By adjusting the template sequences and structures, we found that the catalytic activity of the four DNA homopolymer templates follows the trend: PolyC > PolyA > PolyG > PolyT, indicating that the C base has the strongest affinity for Au and Pt ions. For the same nucleation sequence, the catalytic activity of DNA-Au/Pt nanozymes is closely related to the length and structure of the DNA template sequence used. Based on these findings, combined with the cleavage action of DNAzymes to trigger the transformation of DNA structure, we have proposed a biosensing method for detecting miRNA. Besides, The DNA-Au/Pt nanozyme possesses dual enzyme-like activities of catalase (CAT) and laccase (LAC). Based on the dual-enzyme cascade catalytic system, it can efficiently degrade the organic pollutant malachite green (MG) into less toxic products. This research offers valuable insights into the synthesis of DNA templated nanozymes, and it highlights the potential for optimizing their performance by modulating the DNA sequences. More importantly, this research provides new insights for the design and application of multienzyme activities and multifunctional applications nanozymes in the future.

Fluorophores with color-shifting characteristics have attracted enormous research interest in the quantitative application of RNA sensors. It reports here a simple synthesis, luminescent properties, and co-transcription ability of de-conjugated triphenylmethane leucomalachite green (LMG). This novel clusteroluminescence fluorophore is rapidly synthesized from malachite green (MG) in reductive transcription system containing dithiothreitol, emitting fluorescence in the UV region through space conjugation. The co-transcribed MG RNA aptamer (MGA) bound to the ligand, resulting in red fluorescence from the through-bond conjugation. Given the equilibrated color-shifting fluorophores, they are rationally employed in a 3WJ-based rolling circle transcription switch, with the target-aptamer acting as an activator to achieve steric allosterism. This one-pot system allows the target to compete continuously for allosteric sites, and the activated transcription switches continue to amplify MGA forward, achieving accurate Aflatoxin 1 quantification at the picomolar level in 1 h. Due to the programmability of this RNA sensor, the design method of target-competitive aptamers is standardized, making it universally applicable. A simple strategy for deconjugating malachite green (MG) ligand in a reductive enzymatic environment is reported to manipulate the clusteroluminescence within isolated benzene rings. This strategy is then employed to construct a one-pot, ratiometric RNA sensor using an allosteric transcription-switch and rolling circle transcription. The RNA aptamer (MGA)-MG fluoresces red, while the reductive leucomalachite green (LMG) fluoresces in the UV region. image

Although nanozymes have shown significant potential in wastewater treatment, enhancing their degradation performance remains challenging. Herein, a novel catalytic behavior was revealed for defective nanozymes with catalase-mimicking characteristics that efficiently degraded tetracycline (TC) in wastewater. Hydroxyl groups adsorbed on defect sites facilitated the in-situ formation of vacancies during catalysis, thereby replenishing active sites. Additionally, electron transfer considerably enhanced the catalytic reaction. Consequently, numerous reactive oxygen species (ROS) were generated through these processes and subsequent radical reactions. The defective nanozymes, with their unique catalytic behavior, proved effective for the catalytic degradation of TC. Experimental results demonstrate that •OH, •O2-, 1O2 and e- were the primary contributors to the degradation process. In real wastewater samples, the normalized degradation rate constant for defective nanozymes reached 26.0 min-1 g-1 L, exceeding those of other catalysts. This study reveals the new catalytic behavior of defective nanozymes and provides an effective advanced oxidation process for the degradation of organic pollutants.

The light-up aptamer-dimethylindole red (DIR) complexes have been applied in biochemistry analysis as promising signal transduction tools. However, the unfavorable repulsions between DIR and the long-sequence aptamer switch hinder the complex's further development, and it is urgent to engineer a feasible and efficient strategy for synchronously and rationally adjusting the DIR chemical structure and the DIR aptamer performance. Herein, we communicate a versatile docking-guided rational tailoring strategy to effectively upgrade a DNA aptamer which specifically turns on the fluorescence of a synthesized amino-functionalized DIR analogue (NH2-DIR). After optimizing with three-level tailoring strategies including molecule docking-guided tailoring, coarse tailoring, and fine tailoring, the NH2-DIR aptamer switch with higher binding affinity and specificity, considerable fluorescence-activation ability, and 40% shortened length was obtained. Integrating the experimental and docking results, the binding mechanism between NH2-DIR and the tailored aptamer was deciphered via three types of interactions.

As the up-and-comer in the development of RNA nanotechnology, RNA nanomaterials based on functionalized rolling circle transcription (RCT) have become promising carriers for drug production and delivery. This is due to RCT technology can self-produce polyvalent tandem nucleic acid prodrugs for intervention in intracellular gene expression and protein production. RNA component strands participating in de novo assembly enable RCT-based nanomaterials to exhibit good mechanical properties, biostability, and biocompatibility as delivery carriers. The biostability makes it to suitable for thermodynamically/kinetically favorable assembly, enzyme resistance and efficient expression in vivo. Controllable RCT system combined with polymers enables customizable and adjustable size, shape, structure, and stoichiometry of RNA building materials, which provide groundwork for the delivery of advanced drugs. Here, we review the assembly strategies and the dynamic regulation of RCT-based nanomaterials, summarize its functional properties referring to the bottom-up design philosophy, and describe its advancements in tumor gene therapy, synergistic chemotherapy, and immunotherapy. Last, we elaborate on the unique and practical value of RCT-based nanomaterials, namely "self-production and self-sale", and their potential challenges in nanotechnology, material science and biomedicine.

Nucleic acid-based hydrogels that integrate intrinsic biological properties of nucleic acids and mechanical behavior of their advanced assemblies are appealing bioanalysis and biomedical studies for the development of new-generation smart biomaterials. It is inseparable from development and incorporation of novel structural and functional units. This review highlights different functional units of nucleic acids, polymers, and novel nanomaterials in the order of structures, properties, and functions, and their assembly strategies for the fabrication of nucleic acid-based hydrogels. Also, recent advances in the design of multifunctional and stimuli-responsive nucleic acid-based hydrogels in bioanalysis and biomedical science are discussed, focusing on the applications of customized hydrogels for emerging directions, including 3D cell cultivation and 3D bioprinting. Finally, the key challenge and future perspectives are outlined.

Rolling circle amplification (RCA) technology provides a better alternative for the highly efficient, visual, and rapid on-site detection and poisoning mechanism screening for food-borne microorganisms. The review focuses on the innovative analytical strategies of six primary kinds of microbial targets, including one nucleic acid target and five non-nucleic acid targets (pathogens, proteins, lipopolysaccharides, mycotoxins, and bacterial toxins), covering four recognition elements of antibody, aptamer(Apt), probe and primer, and introduces the latest analysis strategies. These innovative sensing strategies are summarized from five perspectives for the first time, including the combination of RCA technology and immune reactions, functional nucleic acids (FNAs), nanomaterials, multiple amplification technologies, and portable devices, fully demonstrating novel RCA-based biosensors with superior specificity, efficiency, sensitivity, and portability for the detection of food-borne microorganisms. In the meanwhile, the emerging RCA biosensing technology, as a future development prospect of RCA, is proposed as well. This review provides a comprehensive reference for the fields of biomolecular science, disease diagnosis, and health care.

硫酸软骨素(chondroitin sulfate,CS)是一类从动物组织中提取的蛋白类黏性多聚糖,在人体免疫调节、缓解关节疼痛、抗凝活性和保护神经元等方面发挥效用,开发硫酸软骨素传感器对生物医药行业研发相关药剂、保健产品质量的粗筛具有重要意义.利用荧光传感器制备简便、响应迅速、选择性强的特点,用阳离子聚合物PDDA诱导芘基分子HPTS自组装,导致荧光淬灭构建传感器.研究结果显示,待测物的加入引起510 nm处荧光强度的恢复,最终建立了实际样品中CS的快速分析方法.构建的硫酸软骨素荧光传感器在浓度0～5μmol·L-1间呈现良好的线性关系,线性方程为y=1.90752+0.27359x(R2=0.9982),检出限为2.8μmol·L-1,样品加标回收率在104％～121％之间.该传感器具有水溶性好、特异性强、直观可视等特点.

DNA水凝胶作为一种生物合成分子,既具有DNA分子的特异性、生物可降解性和分子识别等特性,又具有水凝胶的高亲水性等特征.刺激响应型DNA水凝胶主要是在环境因素的刺激下,利用常规DNA序列经Watson-Crick碱基互补配对形成的DNA分支结构或多种功能核酸的特殊DNA序列形成的i-motif结构;T-A·T三螺旋结构、C-G· C+三螺旋结构及G-四链体结构等对环境的响应行为使水凝胶形成及应用.近年来,刺激响应型DNA水凝胶因其在温度、pH、光、金属离子、生物分子等单刺激因素,以及光热、金属离子、有机物、温度与pH等多刺激因素下的独特应答性质,在生物传感、生物成像、药物递送、生物材料等方面得到了广泛的应用.综述了刺激响应型DNA水凝胶的形成方法、分类及其核酸来源,形成后的表征手段以及在环境刺激下的响应行为与应用,概括了目前刺激响应型DNA水凝胶的研究热点,并就其未来发展趋势做出了预测.

功能核酸DNA水凝胶是一种以DNA为构建单元通过化学反应或物理缠结自组装而成的新型柔性材料,其构建单元中包含1种或多种能够形成功能核酸的特定序列.功能核酸是通过碱基修饰和DNA分子之间的相互作用力组合的一类特定核酸结构,包括核酸适配体、DNA核酶、G-四联体(G-quadruplex,G4)和i-motif结构等.传统上,高浓度的长DNA链是制备DNA水凝胶的必要条件,而核酸扩增方法的引入为DNA水凝胶的组装方式提供了新的可能.因此,对常用于制备DNA水凝胶的多种功能核酸以及核酸的提取、合成和扩增手段进行了详细的介绍.在此基础上,综述了通过化学或物理交联方式组装功能核酸DNA水凝胶的制备方法.最后,提出了DNA纳米材料的组装所面临的挑战和潜在的发展方向,以期为开发高效组装的功能核酸DNA水凝胶提供参考.

自DNA被开发为纳米级的自组装材料以来,凭借其可调节的多功能性、便利的可编程性、精确的分子识别能力、高通用性以及优越的生物相容性和生物降解性连接了生命科学和材料科学两大领域.受益于DNA纳米材料的结构特性,以功能核酸作为构建单元,经交联、自组装形成的功能核酸DNA水凝胶已成为新型材料领域的研究热点.基于此,对功能核酸DNA水凝胶所具有的主要理化特性进行了总结,并进一步综述了近年来功能核酸DNA水凝胶在药物递送与靶向治疗、生物传感、三维组织构建等生物医学、分子检测及环境工程等领域中的应用进展.最后,从生命科学和材料科学的角度总结了在设计与搭建功能核酸DNA水凝胶时应考虑的关键点和方向,以期助力DNA水凝胶在多学科领域的研究与应用.