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COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS · 2025 · IF 5.4

Construction of a purine base electrochemical sensing platform based on NH2-MXene@COFTA-DH/GCE for simultaneous detection of guanine and adenine

Ruixue Yan,Hao Guo,Zeyun Yang,Wenting Niu,Jianpeng Ma,Liping Peng,Mingyue Wang,Wu Yang

Alterations in guanine (GA) and adenine (AE) levels within the human body are intricately linked to overall health and daily physiological functions. Therefore, it is crucial to develop an accurate and sensitive sensor for quantifying the levels of GA and AE. In this present paper, a novel electrochemical sensor NH 2 -MXene@COF TA-DH /GCE was constructed by in-situ growing COF TA-DH with high crystallinity, good thermal stability and N-rich active centers based on Schiff base condensation reaction of 1, 3, 5-tris-(4-aminophenyl) triazine (TAPT) and 2,5-dihydroxyl-terephthalaldehyde (DHTA) on highly conductive NH 2 -MXene and then modifying it on the surface of the glass carbon electrode (GCE). The highly dispersed sandwich structure of NH 2 -MXene@COF TA-DH significantly increases the specific surface area and exposes a greater number of active sites. Benefitted from remarkable synergistic effect, the sensor exhibited excellent electrochemical catalytic activity and high sensitivity for GA and AE detection. The sensing platform was successfully used for simultaneous detection of GA and AE with wide linear ranges of 0.6–230 μM and 1–450 μM and low detection limits of 0.07 μM and 0.44 μM, respectively. This study expands applications of COFs and MXene in electrochemical biosensing platform and provides a general and valuable strategy for COFs-based electrochemical sensor construction for biomedical investigations

研究领域: 生物医学 · 分析化学
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STAR Protocols · 2022 · IF 0

Protocol for targeted modification of the rice genome using base editing

Meixia Wang,Fang Yan,Huanbin Zhou

Summary Base editing is a precision genome-editing approach that is widely utilized to generate single-nucleotide variants (SNVs) in genomes. Here, we present a protocol to perform targeted adenine (A)-to-guanine (G) substitution in rice using adenine base editor (ABE). We detail the design of sgRNA, CRISPR plasmid construction, rapid genetic transformation of rice, and genotyping of editing events. This protocol can be applied to cytosine base editing in rice as well. For complete details on the use and execution of this protocol, please refer to Yan et al. (2021). 1

研究领域: 植物生物学 · 基因组编辑 · 分子遗传学
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MICROCHEMICAL JOURNAL · 2024 · IF 4.9

Electrochemical voltammetry assessment of cellular DNA damage caused by drugs with different mechanisms: A comparative analysis with the γH2AX focus and alkaline comet assays

Chaoqun Fei,Shulan Qi,Jiahuan Zhang,Shi Zhou,Yingxue Guo,Yanli Zhao,Jiazheng Jin,Jinlian Li,Ximing Cui,Dongmei Wu

As industrialization accelerates, there is an increasing need to find a quick, easy, and affordable method to assess the potential DNA damage caused by newly developed chemicals. Electrochemical voltammetry using intracellular purine bases as markers shows promise for detecting DNA damage. However, it is unclear whether drugs with different DNA damage mechanisms will impact purine metabolism in cells and consequently disturb the electrochemical indicators of DNA damage. In this paper, the electrochemical behaviors of HepG2 cells exposed to the direct DNA-damaging agent (MNNG), the indirect DNA-damaging agent (CPT11), and the non-DNA-damaging agent (KCl) were studied. The findings showed that the electrochemical signals of cells increased with the rising concentration of DNA-damaging drugs. A time-dependent effect was observed, with a lower threshold dose for 4 h compared to 1 h. However, the non-DNA-damaging agent did not induce an increase in the cellular electrochemical signal. Despite the higher cytotoxicity of CPT11, MNNG triggered DNA damage at a lower dose, as detected by electrochemical indicators. The results of the electrochemical indicators assessment of DNA damage correlated significantly with those of the γH2AX focus and alkaline comet assays, with r > 0.98 and P < 0.01. Particularly, there was a stronger correlation with the alkaline comet assay. Furthermore, electrochemical voltammetry detected DNA damage at a lower threshold compared to other assessment methods, demonstrating a higher sensitivity to DNA damage. These findings suggest that by using purine bases as markers, electrochemical voltammetry enables precise assessment of DNA damage caused by various mechanisms.

CAS / MDL: 7447-40-7 70-25-7
研究领域: 毒理学 · 药理学 · 分子生物学 · 生物医学工程
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MICROCHEMICAL JOURNAL · 2020 · IF 3.594

Fluorescent sensing of mercury (II) and copper (II) ions based on DNA-templated Cu/Ag nanoclusters

Xiu-Fang Huang,Bing-Xue Ren,Chi-Fang Peng,Xin-Lin Wei,Zheng-Jun Xie

In this study, the fluorescence enhancement effect of guanine base on DNA-Cu/AgNCs was reported and fluorescent methods for the determination of Hg 2+ or Cu 2+ ion were established. The effect of adjacent and inserted guanine base (G) sequences in DNA template on the fluorescence intensity of DNA-Cu/AgNCs was investigated. It was found that G 5 sequence extended from the 5′ end of a typical C-riched DNA strand, 5′-CCCTTAATCCCC-3′ (DNA1), demonstrated fluorescence enhancement effect in the preparation of the DNA-Cu/AgNCs. Moreover, this G5-DNA-Cu/AgNCs can be used to determine Hg 2+ and Cu 2+ ions due to fluorescence quenching by the two metal ions while the DNA1-Cu/AgNCs only can determine Hg 2+ ions. Specific assays for Hg 2+ ions and Cu 2+ ions were established through masking Cu 2+ ions by EDTA and Hg 2+ ions by NaBH 4 , respectively. In Hg 2+ determination, a linear range from 5.0 nM to 40 nM was obtained with a detection limit of 1.0 nM. In Cu 2+ determination, a wide linear range from 0.01 µM to 5.0 µM was obtained with a detection limit of 5.0 nM. The recoveries of 89.6–106.8% for Cu 2+ ions and 90.6–106.2% for Hg 2+ ions were obtained in the measurements of Hg 2+ and Cu 2+ ions spiked lake samples, respectively. The results indicate the potential application of the DNA-Cu/AgNCs as a metal ions probes in real samples.

CAS / MDL: 16940-66-2 7761-88-8
研究领域: 分析化学 · 环境科学 · 纳米技术 · 生物化学
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BIOMATERIALS · 2023 · IF 14

Photoactivatable base editors for spatiotemporally controlled genome editing in vivo

Quan Zou,Yi Lu,Bo Qing,Na Li,Ting Zhou,Jinbin Pan,Xuejun Zhang,Xuening Zhang,Yupeng Chen,Shao-Kai Sun

CRISPR-based base editors (BEs) are powerful tools for precise nucleotide substitution in a wide range of organisms, but spatiotemporal control of base editing remains a daunting challenge. Herein, we develop a photoactivatable base editor (Mag-ABE) for spatiotemporally controlled genome editing in vivo for the first time. The base editing activity of Mag-ABE can be activated by blue light for spatiotemporal regulation of both EGFP reporter gene and various endogenous genes editing. Meanwhile, the Mag-ABE prefers to edit A4 and A5 positions rather than to edit A6 position, showing the potential to decrease bystander editing of traditional adenine base editors. After integration with upconversion nanoparticles as a light transducer, the Mag-ABE is further applied for near-infrared (NIR) light-activated base editing of liver in transgenic reporter mice successfully. This study opens a promising way to improve the operability, safety, and precision of base editing.

研究领域: 遗传学 · 分子生物学 · 生物技术
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