CHEMOMETRICS AND INTELLIGENT LABORATORY SYSTEMS
·
2015
·
IF 2.321
Jing-Jing Kong,Zeng-Ping Chen,Yao Chen,Xiu-Fang Yan,Ru-Qin Yu
Accurate quantification of genomic DNA is fundamental to many molecular analyses such as diagnostic assays and genotyping. Due to relatively low and varying extraction rates of DNA from cell lysates by genomic DNA extraction kits, it is desired to quantitatively determine DNA contents directly in cell lysates. However, the coexistence of scatterers (e.g., cell debris), absorbers, and intrinsic fluorophores in cell lysates prevents either UV–Vis spectrophotometric methods or fluorometric methods based on fluorescent dyes and ratiometric models to be directly applied to cell lysates. In this contribution, a novel fluorometric method based on an advanced chemometric model which can effectively eliminate the effects of scatterers and absorbers on fluorescence spectra of cell lysates was proposed for the direct quantification of DNA in lysates of CEM cells. Without cumbersome prior DNA extraction, the proposed method achieved quite stable and accurate DNA quantification with recovery rates in the range of 89%–115%, considerably better than that of either the fluorometric method based on the ratiometric model or the UV–Vis spectrophotometric method coupled with DNA extraction. The proposed method has the advantages of rapidness, simplicity, and high precision, and can be developed into a promising alternative for accurate quantification of genomic DNA.
研究领域:
分子生物学
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生物化学
·
分析化学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Fan Yang,JinBo Gong,Ming Li,Xiangyang Jiang,Jiawen Zhang,Meiyan Liao,Hanfei Zhang,Pier-Luc Tremblay,Tian Zhang
PCR-based techniques routinely employed for the detection of mutated linear DNA molecules, including circulating tumor DNA (ctDNA), require large nucleotide sections on both sides of the mutation for primer annealing. This means that DNA fragments with a mutation positioned closer to the extremities are unlikely to be detected. Thus, sensors capable of recognizing linear DNA with characteristic mutations closer to the ends would be advantageous over the state-of-the-art approaches. Here, an electrochemiluminescence-resonance energy transfer (ECL-RET) biosensor comprising capped CdS quantum dots and hairpin DNA probes labeled with Au nanoparticles was developed for the detection of epidermal growth factor receptor (EGFR) ctDNA carrying the critical T790M lung cancer mutation. The ECL-RET system detected different DNA molecules including single-stranded 18-nucleotides (nt) and 40-nt as well as double-stranded 100-nt with the single nucleotide polymorphism (SNP) coding for T790M located either in the middle or only 7 nt from one end. For all target DNA, the sensor’s limits of detection (LODs) were in the aM range, with excellent selectivity. It was the case of 100-nt target linear ctDNA fragments with LODs of 8.1 and 3.4 aM when the EGFR T790M SNP was either in the middle or at the end, respectively. These results show that ECL-RET systems can sense mutations in DNA fragments that would remain undetected by standard techniques.
研究领域:
生物传感器
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纳米技术
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癌症研究
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Miao He,Yan Chen,Feifan Yin,Wenting Cheng,Zhongyun Wang,Yang Xiang
Type 2 diabetes is on the rise year by year, and 80 % of patients with type 2 diabetes die from cardiovascular complications. Strict control of blood glucose can significantly reduce the occurrence and development of diabetic microvascular complications, but can not effectively prevent the occurrence of diabetic macrovascular lesions. DNA methyltransferase 1 (DNMT1) plays an important role in developing phenotypic switching and secretion of inflammatory factors in vascular smooth muscle cells(VSMC), leading to the deregulation of vascular function. In this work, a biosensor based on methylation protection was designed. In the presence of the target DNMT 1, the formed methylation sites protect the dsDNA from being digested by the Hpa II restriction enzyme. At the same time, a large number of G-quadruplex are produced by the catalyzed hairpin assembly (CHA) reaction cycle, and the peroxidase-like activity of G-tetraplex and hemin is used to continuously catalyze the color change of substrate ABTS, thus realizing dual signal amplification. The binding of magnetic beads to dsDNA effectively reduces the background signal and enables a highly selective and sensitive analysis of the target DNMT 1 in complex biological samples, with a detection limit as low as 0.22 UmL −1 , showing a good linear relationship in the range of 0.22–10 UmL −1 . A new approach for the study of macrovascular damage is provided in type 2 diabetes.
研究领域:
医学
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分子生物学
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DOI
高影响因子
Science of Advanced Materials
·
2022
·Vol. 14
·期 5
·911-919
·
IF 1.067
Zhu Chunxia,Ming Shenqiu,Qiu Bin,Zhong Hui
FePt nanoparticles were in this study prepared at varied concentrations to assess their mechanism in regulating DNA injury and inactivation of Hela cells through mitogen-activated protein kinase (MAPK) signal pathway. The cells were divided into following 5 group sets: 1st, 2nd, 3rd,
4th and 5th, which were randomly intervened by following varied concentrations of FePt nanoparticles; 0, 5, 10, 15 and 20 μg/mL. Their action on cellular toxicity, proliferation and inactivation of cells was detected with CCK-8 method, while apoptotic condition was detected with
flow cytometry (FCM). The degree of DNA double chains fracture was detected with neutral single cell gel electrophoresis and immunofluorescence staining. The protein expressions of of EKR1/2, p-ERK1/2, p38 MAPK and p-p38 MAPK in every set were detected with Western Blot assay, while proliferative
activity in the 4th and 5th sets was lower than in the 1st set, and inhibiting action in the 4th set was best. There was no notable difference between 4th and 5th sets, and apoptotic rate in the 1st set was lower than in the other sets. The DNA percentage in the Hela cells was reduced and
Tial length was lengthened along with increased concentration of adopted FePt nanoparticles. The proliferation of Hela cells was restrained by FePt nanoparticles through restraining of the MAPK signal pathway, but growth of Hela cells was not affected, and DNA double-strand break (DSB) was
increased. The FePt nanoparticles could therefore be adopted as one kind of effective anti-tumor drug through increased DNA DSB in tumor cells based on killing effect of tumor cells and good compatibility for normal cells.
研究领域:
纳米技术
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癌症生物学
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细胞信号传导
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毒理学
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DOI
高影响因子
Matter
·
2025
·
IF 17.5
Yakun Gao,Biao Ma,Gangsheng Chen,Chengtao Xu,Ziyan Kong,Yanjie Chen,Chao Zhao,Duxin Chen,Wenwu Yu,Hong Liu
DNA data storage has emerged as a promising solution to address the challenges of data explosion due to its high storage density and durability. However, traditional storage architectures require substrate replacement and the use of environmentally unfriendly chemicals for multistep operations, hindering seamless and sustainable storage. Here, we report liquid metal (LM)-DNA, an integrated and sustainable DNA storage architecture that leverages a responsive liquid metal as a functionality-adaptive storage medium. LM-DNA allows cascaded storage operations within a single medium, including enzymatic DNA synthesis, hermetic encapsulation through thermal activation, and electrokinetic decapsulation driven by the electrocapillary effect. The green enzymatic DNA data writing and physical data encapsulation/decapsulation avoid toxic chemicals, ensuring a more sustainable route for DNA storage. Furthermore, the fluidity of liquid metal enables biomimetic operations such as file splitting, merging, and reshaping, offering unprecedented flexibility in information management. LM-DNA provides a promising foundation for advancing sustainable and biomimetic information storage technologies.
研究领域:
生物技术
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材料科学
·
数据存储
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产品关联
DOI
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
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2015
·
IF 2.729
Shuang Ren,Huan Wang,Hongyi Zhang,Liqing Yu,Mingjie Li,Meng Li
Novel mesoporous carbon fibers (MCFs) were fabricated through a self-templating strategy and solution growth process. MCF modified glassy carbon electrode (MCF/GCE) was proposed for the direct electrocatalytic and simultaneous determination of four DNA bases (guanine (G), adenine (A), thymine (T) and cytosine (C)) using differential pulse voltammetry (DPV) measurement. The MCF/GCE exhibits satisfactory electrocatalytic activity by significantly enhancing the oxidation peak currents and lowering the oxidation potentials toward the four bases when compared with those on GCE. Especially, MCF greatly accelerates the electron transfer of pyrimidine base, which have high oxidation potentials and slow electron transfer kinetics. Moreover, MCF/GCE realizes the simultaneous determination of four DNA bases with low detection limits, high sensitivity and wide linear range. More important, the proposed electrode was successfully applied in the direct detection of bases in herring sperm DNA sample with satisfactory results. Besides, the MCF/GCE shows good long-term stability and reproducibility. These superior performances of MCF/GCE toward purine and pyrimidine DNA bases provide a good possibility for applying the MCF/GCE in our further DNA biosensor research.
研究领域:
生物化学
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分析化学
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DOI
高影响因子
TALANTA
·
2017
·
IF 4.162
Fangfang Li,Xuan Hu,Fengyi Wang,Baozhan Zheng,Juan Du,Dan Xiao
A simple, rapid, and ultrasensitive fluorescence strategy for adenosine triphosphate (ATP) detection was developed by using a FAM (carboxyfluorescein) labeled DNA (FAM-DNA). In this strategy, highly fluorescent FAM-DNA was used as a probe, and nanoceria (CeO 2 NPs) acted as an efficient quencher. FAM-DNA attached to the surface of nanoceria through the coordination between the phosphate group of DNA and NP surface, which induced complete quenching in the FAM-DNA fluorescence due to a photo induced electron transfer (PET) process. It was found that ATP can readily displace adsorbed DNA from nanoceria surface because of the stronger coordination ability of ATP with nanoceria, and the nanoceria-based competitive binding resulted in over 7-fold fluorescence enhancement. Over a wide range from 0.1 nM to 1.5 μM, a good linear relationship between the fluorescence intensity and the concentration of ATP was obtained and the detection limit was estimated to be as low as 54 pM. This method was successfully used to analyze ATP in a single drop of blood and human urine.
研究领域:
生物化学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
ARCHIVES OF TOXICOLOGY
·
2022
·Vol. 97
·期 2
·441-456
·
IF 6.168
Xu Chi,Jiang Shuyun,Ma Xingyu,Jiang Zeyong,Pan Youguang,Li Xiang,Zhang Liying,Zhou Hao,Chen Shen,Xing Xiumei,Chen Liping,Fu Wenfan,Wang Qing,Chen Wen,Li Daochuan
Cisplatin is recommended as a first-line chemotherapeutic agent against advanced non-small cell lung cancer (NSCLC), but acquired resistance substantially limits its clinical efficacy. Recently, DNA methylation has been identified as an essential contributor to chemoresistance. However, the precise DNA methylation regulatory mechanism of cisplatin resistance remains unclear. Here, we found that nicotinamide nucleotide transhydrogenase ( NNT ) was silenced by DNA hypermethylation in cisplatin resistance A549 (A549/DDP) cells. Also, the DNA hypermethylation of NNT was positively correlated to poor prognosis in NSCLC patients. Overexpression of NNT in A549/DDP cells could reduce their cisplatin resistance, and also suppressed their tumor malignancy such as cell proliferation and clone formation. However, NNT enhanced sensitivity of A549/DDP cells to cisplatin had little to do with its function in mediating NADPH and ROS level, but was mainly because NNT could inhibit protective autophagy in A549/DDP cells. Further investigation revealed that NNT could decrease NAD + level, thereby inactivate SIRT1 and block the autophagy pathway, while re-activation of SIRT1 through NAD + precursor supplementation could antagonize this effect. In addition, targeted demethylation of NNT CpG island via CRISPR/dCas9-Tet1 system significantly reduced its DNA methylation level and inhibited the autophagy and cisplatin resistance in A549/DDP cells. Thus, our study found a novel chemoresistance target gene NNT , which played important roles in cisplatin resistance of lung cancer cells. Our findings also suggested that CRISPR-based DNA methylation editing of NNT could be a potential therapeutics method in cisplatin resistance of lung cancer.
研究领域:
肿瘤学
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分子生物学
·
表观遗传学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Ping Xu,Libo Yuan,Ke Wang,Boyuan Pan,Yong Ye,Kui Lu
Two peptide-carbazole conjugates , CTAT and CNLS, were designed and synthesized using carbazole Schiff base to modify the cell membrane penetrating peptide TAT (47–57) and the nuclear localization peptide NLS at the N terminus . The interaction with ctDNA was investigated by multispectral and agarose gel electrophoresis . And the effects of CNLS and CTAT on the G-quadruplex structure were explored by circular dichroism titration experiments. The results show that both CTAT and CNLS interact with ctDNA in a minor groove binding manner. Both conjugates bind more tightly to DNA than the individual substances CIBA, TAT and NLS. In addition, CTAT and CNLS are capable of unfolding parallel G-quadruplex structures and are potential G-quadruplex unfolding agents. Finally, broth microdilution was performed to test the antimicrobial activity of the peptides. The results showed that CTAT and CNLS had a 4-fold increase in antimicrobial activity compared with the parent peptides TAT and NLS. They could exert antimicrobial activity by disrupting the integrity of cell membrane bilayer and binding to DNA, and could be used as novel antimicrobial peptides for the development of novel antimicrobial antibiotics.
研究领域:
生物化学
·
分子生物学
·
药理学
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DOI
PubMed
高影响因子
Epigenomics
·
2017
·
IF 4.541
Hanze Wang,Huican Ke,Yansong Zheng,Junzhong Lai,Qianping Luo,Qi Chen
Aim: Our purpose is to improve the conventional procedures for bisulfite conversion used to detect 5-methylcytosine in DNA. Methods: Impacts of different bisulfite salts, bisulfite conversion temperature, antioxidants and denaturants on DNA conversion and degradation were assessed by methylation-sensitive melt curve analysis. The modified method was tested on different genes and the conversion efficiency was analyzed by bisulfite sequencing. Results: We developed a modified bisulfite conversion method that completes this process within 2 h. We demonstrate that high temperature denaturation is the major cause for DNA degradation, and the addition of ethylene glycol dimethyl ether is an effective way to accelerate the bisulfite conversion. The conversion efficiency is comparable to many other commercial kits. Conclusion: Our modified bisulfite conversion method is simple, cost efficient and less time consuming and is compatible with different genes and samples, thus has a great potential for the future research and clinical applications.
研究领域:
分子生物学
·
表观遗传学
·
基因组学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2011
·
IF 4.311
Lizhu Yang,Ying Xu,Xiuhua Wang,Jing Zhu,Renyi Zhang,Pingang He,Yuzhi Fang
We functionalized aligned carbon nanotubes (ACNTs) electrode with a new kind of β-cyclodextrin (β-CD) derivative through diazotization reaction. The resulting β-CD/ACNTs electrode was used to detect DNA hybridization in homogeneous solution based on host–guest recognition technology . In the sensing protocol, one special DNA probe was designed with a stem-loop structure and both ends modified, which we called dually labeled DNA probe (DLP). One end of the DLP was labeled with dabcyl as guest molecule for β-CD/ACNTs electrode capture, and the other end was labeled with a CdS nanoparticle as an electrochemical tag to indicate the occurrence of DNA hybridization. In the absence of the target DNA sequence , the DLP maintains its hairpin structure in solution phase and would not be captured and detected by the β-CD/ACNTs electrode. In the presence of the complementary target sequence, the conformational structure of the DLP was altered and a double-stranded DNA (dsDNA) molecule was formed by the hybridization of DLP and complementary DNA sequence. Consequently, the dsDNA was captured by the β-CD/ACNTs electrode owing to guest–host recognition between β-CD and dabcyl. The electrochemical signal from the CdS nanoparticle–dsDNA/β-CD/ACNTs was then measured. Under optimized detection conditions, the proposed method showed high sensitivity and selectivity with a detection limit of 5.0 × 10 −13 M for complementary DNA sequence.
研究领域:
纳米技术
·
电化学
·
生物传感器
·
分子识别
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2013
·
IF 5.695
Chun Yang,Lei Liu,Tao Zeng,Daowu Yang,Zhiyi Yao,Yuliang Zhao,Hai-Chen Wu
Both Pb2+ and Ba2+ can bind with high affinity to some specific DNA sequences, inducing the formation of unimolecular G-quadruplex structures. Translocation of a DNA probe containing such sequences through an α-hemolysin nanopore in the presence of Pb2+ or Ba2+ would result in much prolonged DNA translocation events. Quantification of these events can reveal the concentrations of Pb2+ or Ba2+ at as low as 0.8 nM. Besides, Pb2+ and Ba2+ in the solution can be simultaneously detected and individually identified. Furthermore, the probe is highly selective for Pb2+ and Ba2+ detection without interference from other metal ions. This sensing strategy can be extended to many other analytes which can bind to DNA molecules with high affinity.
研究领域:
分析化学
·
纳米技术
·
生物传感器
·
环境科学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）