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PE6c greatly enhances prime editing in transgenic rice plants

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Journal of Integrative Plant Biology 2024年第9期66卷 1864-1870页

作者： Zhenghong Cao Wei Sun Dexin Qiao Junya wang Siyun Li Xiaohan Liu Cuiping Xin Yu Lu Syeda Leeda Gul xue‐chen wang Qi-Jun chen State Key Laboratory of Plant Environmental Resilience College of Biological SciencesChina Agricultural UniversityBeijing100193China Center for Crop Functional Genomics and Molecular Breeding China Agricultural UniversityBeijing 100193China

Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop *** new types of prime editors(PEs)named PE6a–d were recently generated using evolved and engineered reverse transcriptase(RT)variants from three different *** this study,we tested the editing efficiencies of four PE6 variants and two additional PE6 constructs with double-RT modules in transgenic rice(Oryza sativa)***6c,with an evolved and engineered RT variant from the yeast Tf1 retrotransposon,yielded the highest prime-editing *** average fold change in the editing efficiency of PE6c compared with PEmax exceeded 3.5 across 18 agronomically important target sites from 15 *** also demonstrated the feasibility of using two RT modules to improve prime-editing *** results suggest that PE6c or its derivatives would be an excellent choice for prime editing in monocot *** addition,our findings have laid a foundation for prime-editing-based breeding of rice varieties with enhanced agronomically important traits.

关键词： genome editing PE6 PE6c prime editing prime editor rice

Expression of the DNA-repair protein XRCC1 and DNA-repair enzyme Ku70 mRNA in the rat cerebral cortex following global cerebral ischemia/reperfusion Is this correlated with neuroprotective effects of theanine?

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Neural Regeneration Research 2010年第1期5卷 41-46页

作者： Rongliang xue Wenchao chen Ning wang Shasha Li Department of Anesthesiology Second Affiliated Hospital Medical College of Xi'an Jiaotong University Xi'an 710004 Shaanxi ProvinceChina

BACKGROUND： Numerous studies have shown that apoptosis, caused by cerebral ischemia/reperfusion injury, is a dynamic evolving process that occurs in a time-dependent fashion. Non-apoptotic cells can result in apoptosis, with a prolonged reperfusion period and an accumulation of DNA damage. Recent studies have indicated that theanine has a protective effect on the brain. Nonetheless, there is no research relating to whether theanine is neuroprotective following brain injury. OBJECTIVE： To investigate the effects of theanine on apoptosis in the cerebral cortex and mRNA expression of the DNA repair protein, XRCC1, and DNA repair enzyme, Ku70, at various time points following global cerebral ischemia/reperfusion in rats. DESIGN, TIME AND SETTING： A randomized, controlled animal study was performed at the Pharmacological Laboratory of Animal Experimental Center of Xi＇an Jiaotong University, China, from April to October 2007. MATERIALS： Theanine （Wuhan Remote Technology Co., Ltd., China）, XRCC1, Ku70 and GAPDH primers （Shanghai Sangon Biological Engineering Technology ＆ Services Co., Ltd., China） were used in this study. METHODS： A total of 108 healthy, male Sprague Dawley rats were randomly divided into sham operation, ischemia/reperfusion and theanine groups. Each group was further divided into 6 subgroups, with six rats in each, according to reperfusion time （2, 6, 12, 24, 48 and 72 hours）. Rat models of global cerebral ischemia/reperfusion were established by the four-vessel occlusion method. Theanine （1 000 mg/kg） was injected into the caudal vein of rats in the theanine group immediately after surgery. Saline （4 mL/kg） was used in the sham operation and ischemia/reperfusion groups. The same dose was administered every 24 hours. MAIN OUTCOME MEASURES： Flow cytometry was employed to determine apoptosis of cerebral cortical neurons. Real-time reverse transcdption-polymerase chain reaction was applied to quantify the expression of XRCC1 and Ku70 mRNA in the cerebral cor

关键词： brain ischemia/reperfusion injury XRCC1 Ku70 theanine

Excessive Copper Induces the Production of Reactive Oxygen Species,which is Mediated by Phospholipase D, Nicotinamide Adenine Dinucleotide Phosphate Oxidase and Antioxidant Systems

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Journal of Integrative Plant Biology 2008年第2期50卷 157-167页

作者： Zhong-Lian Yu Jin-Guang Zhang xue-chen wang Jia chen State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100094 China

Tobacco BY-2 suspension cells were used to study the chemical damage and its associated mechanisms caused by Cu^2＋. Treatment with 100 μmol/L Cu^2＋ generated a large amount of HzOz and thiobarbituric acid-reactive substances （TBARS） in cells. Using phospholipase D （PLD） specific inhibitor （1-butanol） or phosphatidic acid （PA）, we demonstrated that PLD plays an important role in the generation of H2O2 and TBARS. Semi-quantitative reverse-transcriptase polymerase chain reaction and enzyme activity assays with wild type and nicotinamide adenine dinucleotide phosphate （NADPH） oxidaseoverexpressing BY-2 cells revealed that PLD and PA are the key factors leading to NADPH oxidase activation, which is responsible for H2O2 and TBARS production induced by Cu^2＋. Moreover, the content of ascorbic acid （AsA）, an effective antioxidant, was sharply reduced in BY-2 cells exposed to excessive Cu^2＋. Furthermore, a significant downregulation of the enzymes of AsA biosynthesis and the antioxidant system was found. This evidence suggests that excessive Cu^2＋-elevated reactive oxygen species （ROS） production is caused by upregulated PLD that elevates the activity of NADPH oxidase and its collapsed antioxidant systems that scavenges ROS.

关键词： ascorbic acid Cu^2+ nicotinamide adenine dinucleotide phosphate oxidase phospholipase D reactive oxygen species.

Ethylene Response Factor TERF1 Enhances Glucose Sensitivity in Tobacco through Activating the Expression of Sugar-related Genes

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Journal of Integrative Plant Biology 2009年第2期51卷 184-193页

作者： Ang Li Zhijin Zhang xue-chen wang Rongfeng Huang National Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100094 China Biotechnology Research Institute the Chinese Academy of Agricultural Sciences Beijing 100081 China National Key Facility of Crop Gene Resources and Genetic Improvement Beijing 100081 China National Center for Plant Gene Research (Beijing) Beijing 100081 China

Ethylene response factor （ERF） proteins are important plant-specific transcription factors. Increasing evidence shows that ERF proteins regulate plant pathogen resistance, abiotic stress response and plant development through interaction with different stress responsive pathways. Previously, we revealed that overexpression of TERF1 in tobacco activates a cluster gene expression through interacting with GCC box and dehydration responsive element （DRE）, resulting in enhanced sensitivity to abscisic acid （ABA） and tolerance to drought, and dark green leaves of mature plants, indicating that TERF1 participates in the integration of ethylene and osmotic responses. Here we further report that overexpression of TERF1 confers sugar response in tobacco. Analysis of the novel isolated tomato TERF1 promoter provides information indicating that there are many cis-acting elements, including sugar responsive elements （SURE） and W box, suggesting that TERF1 might be sugar inducible. This prediction is confirmed by results of reverse transcription-polymerase chain reaction amplification, indicating that transcripts of TERF1 are accumulated in tomato seedlings after application of glucose. Further investigation indicates that the expression of TERF1 in tobacco enhances sensitivity to glucose during seed germination, root and seedling development, showing a decrease of the fresh weight and root elongation under glucose treatment. Detailed investigations provide evidence that TERF1 interacts with the sugar responsive cis-acting element SURE and activates the expression of sugar response genes, establishing the transcriptional regulation of TERF1 in sugar response. Therefore, our results deepen our understanding of the glucose response mediated by the ERF protein TERF1 in tobacco.

关键词： Ethylene response factor protein TERF1 glucose sugar responsive gene tobacco

Evaluation of visual quality of spherical and aspherical intraocular lenses by Optical Quality Analysis System

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International Journal of Ophthalmology(English edition) 2017年第6期10卷 914-918页

作者： Yan chen xue wang Chuan-Di Zhou Qiang Wu Department of Ophthalmology Shanghai Jiao Tong University Affiliated First People＇s Hospital Shanghai 200080 China

AIM： To evaluate the impact of spherical and aspherical intraocular lenses on the postoperative visual quality of age-related cataract patients using Optical Quality Analysis System （OQAS）. METHODS： Seventy-four eyes with age-related cataracts were randomly divided into spherical and aspherical lens implantation groups. Best-corrected visual acuity （BCVA） was measured preoperatively, one day, one week, two weeks, one month and two months after surgery. A biometric systems analysis using the OQAS objective scattering index （OSI） was performed. RESULTS： There were no significant differences in visual acuity （P〉0.05） before and after spherical and aspheric lens implantation. There was a negative linear correction between the OSI value and BCVA （t-=-0.634, P=-0.000）, and positive corrections between the OSI value and the lens LOCUS III value of nucleus color （NC）, nucleus opacity （NO）, cortex （C） and posterior lens capsular （P）（r=0.704, P=0.000; r=0.514, P=0.000; r=0.276, P=0.020; r=0.417, P=-0.000, respectively）. OSI values of spherical vs aspherical lenses were 11.5±3.6 vs 11.8±3.4, 4.1±0.9 vs 3.3±0.8, 3.5±0.9 vs 2.7±0.7, 3.3±0.8 vs 2.6±0.7, 3.2±0.7 vs 2.5±0.8, and 3.2±0.8 vs 2.50.8 before and ld, 1, 2wk, 1 and 2mo after surgery, respectively. All time points varied significantly （P〈0.01） between the two groups. CONCLUSION： Aspherical IOLs does not significantly affect visual acuity compared with spherical IOLs. The OSI value, was significantly lower in the aspherical lens group compared with the spherical lens. This study shows that objective visual quality of aspheric IOLs is better than that of the spherical lens by means of OQAS biological measurement method.

关键词： optical quality analysis system age-relatedcataract spherical lens aspherical lens objective scatteringindex

Double Moving Extremes Ranked Set Sampling Design

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Acta Mathematicae Applicatae Sinica 2024年第1期40卷 75-90页

作者： Meng chen wang-xue chen Rui Yang Department of Mathematics and Statistics Jishou UniversityJishou416000China

The traditional simple random sampling(SRS)design method is inefficient in many *** proposed some new designs to increase *** this paper,as a variation of moving extremes ranked set sampling(MERSS),double MERSS(DMERSS)is proposed and its properties for estimating the population mean are *** turns out that,when the underlying distribution is symmetric,DMERSS gives unbiased estimators of the population ***,it is found that DMERSS is more efficient than the SRS and MERSS methods for usual symmetric distributions(normal and uniform).For asymmetric distributions considered in this study,the DMERSS has a small bias and it is more efficient than SRS for usual asymmetric distribution(exponential)for small sample sizes.

关键词： double moving extremes ranked set sampling symmetric distribution efficiency

Interdiffusion behavior between Cr and Zr and its effect on the microcracking behavior in the Cr-coated Zr-4 alloy

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Nuclear Science and Techniques 2021年第12期32卷 1-12页

作者： Ji-Shen Jiang Dong-Qing wang Ming-Yue Du Xian-Feng Ma chen-xue wang Xiu-Jie He Sino-French Institute of Nuclear Engineering and Technology Sun Yat-Sen UniversityZhuhai 519082China

High-temperature chromium(Cr)-zirconium(Zr)interdiffusion commonly occurs in Cr-coated zircaloys applied for enhanced accident-tolerant fuel(ATF)clad*** interdiffusion changes the interfacial microstructure and thus the fracture mechanism of the coating under external *** this study,the interdiffusion behavior in a magnetron sputtered Cr coating deposited on a Zr-4 alloy was studied in a vacuum environment at 1160*** addition,the effect of interdiffusion on the microcracking behavior of the Cr coating was determined by in situ three-point bending *** experimental results show that the interdiffusion behavior resulted in the formation of a ZrCr2 layer,accompanied by the consumption of Cr coating and interfacial *** growth of the diffusion layer followed a nearly parabolic law with respect to annealing time,and the residual stress of the annealed coating decreased with increasing annealing *** external loading,a large number of cracks were generated in the brittle interlayer,and some interfacial cracks were formed and grew at the ZrCr2/Zr-4 *** the remarkable microcracks in the ZrCr2 layer,the vacuum-annealed Cr coating has significantly fewer cracks than the original coating,mainly because of the recrystallization of the coating during annealing.

关键词： Accident-tolerant fuel Surface coating Interdiffusion Three-point bending test Crack propagation

A unique 3D microporous MOF constructed by cross-linking 1D coordination polymer chains for effectively selective separation of CO_(2)/CH4 and C_(2)H_(2)/CH_(4)

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Chinese Chemical Letters 2021年第3期32卷 1153-1156页

作者： Yu-Pei Xia chen-xue wang Mei-Hui Yu Xian-He Bu School of Materials Science and Engineering National Institute for Advanced MaterialsNankai UniversityTianjin 300350China State Key Laboratory of Elemento-Organic Chemistry College of ChemistryNankai UniversityTianjin 300071China

Selective separation of CO_(2)/CH_(4)and C2 H_(2)/CH_(4)are promising for their high-purity industrial demand and scientific research on account of the similar molecular radius and physical *** this work,a unique 3 D microporous MOF material[Cu(SiF6)(sdi)2]·solvents(1,sdi=1,1’-sulfonyldiimidazole)was successfully constructed by cross-linking 1 D coordination polymer *** dense functional active sites on the inner walls of the channel of la can provide strong binding affinities to CO_(2),C2 H_(2),and thus effectively improve the gas separation performance of CO_(2)/CH_(4)and C2 H_(2)/CH_(4).

关键词： Metal-organic framework Cross-linking Selective separation Dense functional active site

Optimized prime editing efficiently generates heritable mutations in maize

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Journal of Integrative Plant Biology 2023年第4期65卷 900-906页

作者： Dexin Qiao Junya wang Min‐Hui Lu Cuiping Xin Yiping Chai Yuanyuan Jiang Wei Sun Zhenghong Cao Siyi Guo xue‐chen wang Qi‐Jun chen State Key Laboratory of Plant Physiology and Biochemistry College of Biological SciencesChina Agricultural UniversityBeijing 100193China Center for Crop Functional Genomics and Molecular Breeding China Agricultural UniversityBeijing 100193China Collaborative Innovation Center of Crop Stress Biology Henan ProvinceInstitute of Plant Stress BiologySchool of Life ScienceHenan UniversityKaifeng 475004China

Low efficiency is the main obstacle to using prime editing in maize(Zea mays).Recently,prime-editing efficiency was greatly improved in mammalian cells and rice(Oryza sativa)plants by engineering primeediting guide RNAs(pegRNAs),optimizing the prime editor(PE)protein,and manipulating cellular determinants of prime *** this study,we tested PEs optimized via these three strategies in *** demonstrated that the ePE5max system,composed of PEmax,epegRNAs(pegRNA-evopreQ.1),nicking single guide RNAs(sgRNAs),and MLH1dn,efficiently generated heritable mutations that conferred resistance to herbicides that inhibit 5-enolpyruvylshikimate-3-phosphate synthase(EPSPS),acetolactate synthase(ALS),or acetyl CoA carboxylase(ACCase)***,we demonstrate that the ePE5max system has sufficient efficiency to generate heritable(homozygous or heterozygous)mutations in maize target genes and that the main obstacle to using PEs in maize has thus been removed.

关键词： CRISPR/Cas epegRNAs herbicide resistance maize MLH1dn prime editing