Foods
·
2026
·Vol. 15
·期 10
·1790
·
IF 6
Xiaorong Xu,Jie Fang,Lingyan Mao,Yingying Wu,Hai Cheng,Jinru Lin,Liyu Shi,Jiali Xing,Xiaohu Luo
DNA must be efficiently extracted from samples to accurately test the authenticity of food, particularly from processed matrices in which DNA integrity may be compromised. We systematically evaluated the efficiency of extracting DNA from dairy and blood products by four methods, namely SDS-CTAB, SDS-isopropanol precipitation, guanidine isothiocyanate magnetic beads, and a commercial kit. The guanidine isothiocyanate-magnetic bead method yields high quantities and purity of DNA; for example, the yield obtained from chicken blood samples was 318.34 ± 4.77 ng/µL, with an A260/A280 ratio ranging from 1.8 to 2.0. The processing time of this method was compared with the DNA Extraction Kit shorter by 40% and unlike methods such as the SDS-CTAB protocol, does not require the use of toxic reagents such as phenol or chloroform, meeting green chemistry requirements. Among the dairy and blood samples tested, it enables the extraction of DNA in quantities comparable to those obtained using commercial kits; moreover, the DNA yield achieved is 20–30% higher than that of these kits. Furthermore, this method is free from the limitations associated with protein contamination and amplification instability often encountered in protocols such as the CTAB-SDS and SDS-isopropanol methods. The magnetic bead approach was adaptable for complex matrices and demonstrated strong tolerance to coexisting contaminants, thereby improving extraction performance in challenging food samples. The magnetic bead surface functionalization and buffer systems could be improved to further increase their versatility. This method enables reliable DNA extraction and advanced technical support for DNA analysis.
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JOURNAL OF ENVIRONMENTAL MANAGEMENT
·
2026
·
IF 9.2
Xiaoying Li,Di Zhang,Jiani Wang,Huang Zhang,Fangfang Li,Yuliang Dong
Environmental DNA (eDNA) has emerged as a global contaminant due to its potential health risks. Although catalytic degradation has been explored for eDNA removal, no study has systematically compared its degradation efficiency and mechanisms under different oxidant systems. This study examined the catalytic degradation of herring sperm DNA (hsDNA) using iron-modified biochar (FBC) activated by persulfate (PS), urea hydrogen peroxide (UHP), and peroxymonosulfate (PMS). The FBC400/PMS system achieved the fastest hsDNA degradation, removing 93.1% of hsDNA within 90 min. Quenching experiments and electron paramagnetic resonance spectroscopy showed that singlet oxygen ( 1 O 2 ) dominated in the FBC400/PMS and FBC400/PS systems, whereas hydroxyl radicals (•OH) dominated in the FBC400/UHP system. Gel electrophoresis and atomic force microscopy confirmed that all systems induced hsDNA strand breakage. The degradation pathways of thymine, a representative DNA base, were further elucidated. In the FBC400/PMS system, 1 O 2 and SO 4 •− synergically attacked thymine C 5 =C 6 , generating dimer and ring-opening products. In the FBC400/UHP system, •OH dominated the competitive pathway of dihydroxylation and methyl oxidation. In the FBC400/PS system, 1 O 2 , SO 4 •− , and •OH synergically drove the C 5 and C 6 hydroxylation and ring opening. These findings offer new insight into eDNA degradation by FBC-activated oxidants and emphasized the importance of selecting an appropriate oxidant system.
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BIOCONJUGATE CHEMISTRY
·
2016
·
IF 4.5
Tieyan Wang,Qixian Chen,Hongguang Lu,Wei Li,Zaifen Li,Jianbiao Ma,Hui Gao
The dilemma of poly(ethylene glycol) surface modification (PEGylation) inspired us to develop an intracellularly sheddable PEG palisade for synthetic delivery systems. Here, we attempted to conjugate PEG to polyethylenimine (PEI) through tandem linkages of disulfide-bridge susceptible to cytoplasmic reduction and an azobenzene/cyclodextrin inclusion complex responsive to external photoirradiation. The subsequent investigations revealed that facile PEG detachment could be achieved in endosomes upon photoirradiation, consequently engendering exposure of membrane-disruptive PEI for facilitated endosome escape. The liberated formulation in the cytosol was further subjected to complete PEG detachment relying on disulfide cleavage in the reductive cytosol, thus accelerating dissociation of electrostatically assembled PEI/DNA polyplex to release DNA by means of polyion exchange reaction with intracellularly charged species, ultimately contributing to efficient gene expression.
研究领域:
基因治疗
·
药物递送系统
·
纳米医学
·
生物材料
·
分子生物学
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ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Xueping Yang,Jingyun Li,Suixin Zhang,Chao Li,Jiehua Ma
CRISPR/Cas-based systems are highly attractive for developing next-generation diagnostic technologies because of their intrinsic merits such as simplicity, sensitivity, and specificity. However, currently, nucleic acid amplification procedures are still needed to achieve attomolar sensitivity in most CRISPR/Cas-based assays, which causes high cost, operation difficulty, and low efficiency. Herein, we combine the CRISPR/Cas12a-based assay and a single-microbead detection platform for one-step and amplification-free detection of DNA at the single-molecule level. By modifying DNA reporters on a biomimetic membrane-coated microbead, the activated Cas12a by targets will cleave these reporters and lighten the bead within 10 min. The method allows the detection of the target down to three copies in a 5 μL sample. Furthermore, we successfully apply this method for the specific identification of viral infection, foodborne bacteria, and DNA mutation in real samples without extra nucleic acid amplification. We believe that this approach offers new insights for developing CRISPR/Cas-based DNA assays in biomedical applications.
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ANALYST
·
2023
·Vol. 148
·期 7
·1492-1499
·
IF 4.2
Jiahai Wang,Lanfang Chen,Cenlin Gui,Jianji Zhu,Baian Zhu,Zhuobin Zhu,Yunhui Li,Daqi Chen
DNA methylation has been considered an essential epigenetic biomarker for diagnosing various diseases, such as cancer. A simple and sensitive way for DNA methylation level detection is necessary. Inspired by the label-free and ultra-high sensitivity of solid-state nanopores to double-stranded DNA (dsDNA), we proposed a nanopore counter for evaluating DNA methylation by integrating a dual-restriction endonuclease digestion strategy coupled with polymerase chain reaction (PCR) amplification. Simultaneous application of BstUI/HhaI endonucleases can ensure the full digestion of the unmethylated target DNA but shows no effect on the methylated ones. Therefore, only the methylated DNA remains intact and can trigger the subsequent PCR reaction, producing a large quantity of fixed-length PCR amplicons, which can be directly detected through glassy nanopores. By simply counting the event rate of the translocation signals, the concentration of methylated DNA can be determined to range from 1 aM to 0.1 nM, with the detection limit as low as 0.61 aM. Moreover, a 0.01% DNA methylation level was successfully distinguished. The strategy of using the nanopore counter for highly sensitive DNA methylation evaluation would be a low-cost but reliable alternative in the analysis of DNA methylation.
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ACS Nano
·
2023
·
IF 17.1
Cuiping Mao,Shuchen Wang,Jiankai Li,Zhuowei Feng,Tong Zhang,Rui Wang,Chunhai Fan,Xingyu Jiang
DNA as an exceptional data storage medium offers high information density. However, DNA storage requires specialized equipment and tightly controlled environments for storage. Fast encapsulation within minutes for enhanced DNA stability to do away with specialized equipment and fast DNA extraction remain a challenge. Here, we report a DNA microlibrary that can be encapsulated by metal–organic frameworks (MOFs) within 10 min and extracted (5 min) in a single microfluidic chip for automated and integrated DNA-based data storage. The DNA microlibrary@MOFs enhances the stability of data-encoded DNA against harsh environments. The encoded information can be read out perfectly after accelerated aging, equivalent to being readable after 10 years of storage at 25 °C, 50% relative humidity, and 10 000 lx sunlight radiation. Moreover, the library enables fast retrieval of target data via flow cytometry and can be reproduced after each access.
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CANCER SCIENCE
·
2023
·
IF 5.7
Bo‐Wen Zuo,Wan‐Xin Yao,Meng‐Die Fang,Juan Ren,Ling‐Lan Tu,Run‐Jie Fan,Yan‐Mei Zhang
The Brother of Regulator of Imprinted Sites (BORIS, gene symbol CTCFL) has previously been shown to promote colorectal cancer cell proliferation, inhibit cancer cell apoptosis, and resist chemotherapy. However, it is unknown whether Boris plays a role in the progression of in situ colorectal cancer. Here Boris knockout (KO) mice were constructed. The function loss of the cloned Boris mutation that was retained in KO mice was verified by testing its activities in colorectal cell lines compared with the Boris wild‐type gene. Boris knockout reduced the incidence and severity of azoxymethane/dextran sulfate‐sodium (AOM/DSS)‐induced colon cancer. The importance of Boris is emphasized in the progression of in situ colorectal cancer. Boris knockout significantly promoted the phosphorylation of γH2AX and the DNA damage in colorectal cancer tissues and suppressed Wnt and MAPK pathways that are responsible for the callback of DNA damage repair. This indicates the strong inhibition of colorectal cancer in Boris KO mice. By considering that the DSS‐promoted inflammation contributes to tumorigenesis, Boris KO mice were also studied in DSS‐induced colitis. Our data showed that Boris knockout alleviated DSS‐induced colitis and that Boris knockdown inhibited the NF‐κB signaling pathway in RAW264.7 cells. Therefore Boris knockout eliminates colorectal cancer generation by inhibiting DNA damage repair in cancer cells and relieving inflammation in macrophages. Our findings demonstrate the importance of Boris in the development of in situ colorectal cancer and provide evidence for the feasibility of colorectal cancer therapy on Boris.
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ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Jingpu Zhang,Lei Pan,Yan Wang,Lin Yin,Li Xu,Jie Tao,Lijun Zhang,Zhaoqin Zhu,Daxiang Cui,Feng Li,Tie Fu Liu
Tryptophan (Trp) as an essential amino acid plays critical roles in regulating multiple cell activities, and the changes of its circulating level usually indicate disease status such as the severity of acute inflammation sepsis. However, the current technology for Trp detection mostly relies on chromatography that cannot meet the rapid and simple detection requirement in monitoring sepsis. Herein, a label-free fluorescent nanosensor was constructed to detect Trp and its carrier protein - human serum albumin (HSA) in plasma. The nanosensor consists of a cytosine (C) - rich signal probe of DNA-templated silver nanoclusters (AgNCs/DNA) and a Trp (Trp) aptamer - based capture probe with a guanine (G) - rich overhang. Trp was found to trigger G-rich sequence-mediated fluorescence enhancement effect for AgNCs/DNA under UV irradiation which offers energy to induce the redox process between limited Trp and silver ions bound on the DNA probes . Based on this photochemical property, the nanosensor exhibited a linear response in the range of 0.05–60 μM with the limit of detection of 0.43 μM for Trp, superior to the current fluorescence-based detection method, and it gave specific response to indole group. When applied in plasma detection, the nanosensor resisted physiological level of NaCl in plasma, but was quenched by trace volume of HSA, which facilitates the combined HSA detection using only 1 μL plasma sample. The simple procedure of “mix, exposure and detection” together with its ultralow sampling volume, time-saving, cost-effective, sensitive and selective properties endow the nanosensor great potentials for future Trp detection-based clinical use.
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MICROCHIMICA ACTA
·
2022
·Vol. 189
·期 6
·1-9
·
IF 6.408
Gao Hongfei,Cui Dandan,Zhai Shanshan,Yang Yao,Wu Yuhua,Yan Xiaohong,Wu Gang
A label-free electrochemical impedimetric biosensor was constructed based on gold carbon dots (GCDs) modified screen-printed carbon electrode for the detection of genetic modified (GM) soybean. The structure and property of GCDs were investigated. The GCDs can directly bind to single-stranded DNA probes through Au-thiol interaction and boost electric conductivity for the DNA sensor construction. The quantification of target DNA was monitored by the change of electron-transfer resistance ( R et ) upon the DNA hybridization on sensor surface. Under the optimal conditions, the R et response (vs. Ag reference electrode) increased with the logarithm of target DNA concentrations in a wide linear range of 1.0 × 10 −7 − 1.0 × 10 −13 M with a detection limit of 3.1 × 10 −14 M ( S / N = 3). It was also demonstrated that the proposed DNA sensor possessed high specificity for discriminating target DNA from mismatched sequences. Moreover, the developed biosensor was applied to detect SHZD32-1 in actual samples, and the results showed a good consistency with those obtained from the gel electrophoresis method. Compared with the previous reports for DNA detection, the label-free biosensor showed a comparatively simple platform due to elimination of complicated DNA labeling. Therefore, the proposed method showed great potential to be an alternative device for simple, sensitive, specific, and portable DNA sensor. Graphical abstract
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Micromachines
·
2022
·Vol. 13
·期 1
·83
·
IF 3.523
Kai Guo,Zirui Song,Gaoxing Wang,Chengchun Tang
Microbial activity has gained attention because of its impact on the environment and the quality of people’s lives. Most of today’s methods, which include genome sequencing and electrochemistry, are costly and difficult to manage. Our group proposed a method using the redox potential change to detect microbial activity, which is rooted in the concept that metabolic activity can change the redox potential of a microbial community. The redox potential change was captured by a biosensor consisting of porous boron nitride, ATP-DNA aptamer, and methylene blue as the fluorophore. This assembly can switch on or off when there is a redox potential change, and this change leads to a fluorescence change that can be examined using a multipurpose microplate reader. The results show that this biosensor can detect microbial community changes when its composition is changed or toxic metals are ingested.Keywords:redox potential;aptamer;dysbiosis;boron nitride;fluorescence
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Journal of Materials Chemistry B
·
2022
·Vol. 10
·期 9
·1386-1392
·
IF 7.571
Ling Yang,Ping’an Ma,Xiaorui Chen,Ziyong Cheng,Jun Lin
Sensitive detection of circulating tumor DNA (ctDNA) in vitro has attracted growing attention owing to its potential application in diagnostics of cancer. In this study, we synthesized hydrophilic AgInS2@ZnS core–shell quantum dot nanocrystals and magnetic Fe3O4 nanoparticles, and then the ctDNA triggered hybridization chain reaction was used to detect the CYFRA21-1 DNA associated with lung cancer. In the presence of CYFRA21-1 DNA, three hairpin structures were activated to turn on successively, resulting in the accumulation of quantum dots and eliciting considerable changes of the fluorescence signal. Compared with the conventional fluorescence detection, Fe3O4 provides magnetic adsorption properties and a large surface area for immobilizing and aggregating quantum dot nanoparticles attached to single-stranded DNA. The concentration of CYFRA21-1 is closely related to the number of quantum dots remaining after magnetic adsorption, which provides a promising approach for ctDNA quantification.
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BIOSENSORS & BIOELECTRONICS
·
2022
·
IF 12.545
Xiaojuan Wang,Zefeng Mao,Ruipeng Chen,Shuning Li,Shuyue Ren,Jun Liang,Zhixian Gao
DNA origami technology has great potential for biosensor applications. Here, we described the construction of a self-assembled DNA origami biosensor for the precise localization of fluorescent aptamers . Due to the molecular weight difference between DNA origami and aptamer, centrifugal filters were used to quantitatively detect adenosine triphosphate (ATP). The ATP-specific aptamer labeled with fluorescence reporter 6-carboxyfluorescein FAM (FAM-aptamer) was selected as the recognition element and signal probe . ATP duplexed aptamers bound to triangular DNA origami by base-complementary pairing, resulting in high fluorescence signals on the origami arrays. The competitive binding of ATP toward the FAM-aptamer triggered the release of FAM-aptamer-ATP complexes from the surface of the origami array, resulting in weakened fluorescence signals. For ATP quantification, 100 kD centrifugal filters were employed, followed by measurement of the fluorescence signal trapped on the origami arrays of the filter device. The successful synthesis of origami-aptamer arrays was characterized by atomic force microscopy , laser confocal microscopy , and electrophoresis . Fluorescence measurements exhibited an excellent linear relationship with logarithms of ATP concentrations within 0.1–100 ng mL −1 , with a detection limit of 0.29 ng mL −1 . By replacing aptamers and complementary strands, we demonstrated the potential of this method for 17β-estradiol detection. Considering that the detection mechanism is based on the hybridization and displacement of DNA strands, the detection system had the potential for recharging. Our study provides new insights into applying DNA origami technology in small molecule detection.
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