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CRISPR/Cas9-based gene activation and base editing in Populus

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Horticulture Research 2023年第6期10卷 205-213页

作者： Tao Yao Guoliang Yuan Haiwei Lu Yang Liu jin zhang Gerald A.Tuskan Wellington Muchero jin-Gui Chen Xiaohan Yang Biosciences Division Oak Ridge National LaboratoryOak RidgeTN 37831USA The Center for Bioenergy Innovation Oak Ridge National LaboratoryOak RidgeTN 37831USA Chemical and Biological Process Development Group Pacific Northwest National Laboratory902 Battelle BoulevardRichlandWA 99352USA Department of Academic Education Central Community College-HastingsHastingsNE 68901USA State Key Laboratory of Subtropical Silviculture College of Forestry and BiotechnologyZhejiang A&F UniversityHangzhou 311300China

The genus Populus has long been used for environmental,agroforestry and industrial applications *** Populus is also recognized as a desirable crop for biofuel production and a model tree for physiological and ecological *** such,various modern biotechnologies,including CRISPR/Cas9-based techniques,have been actively applied to Populus for genetic and genomic improvements for traits such as increased growth rate and tailored lignin ***,CRISPR/Cas9 has been primarily used as the active Cas9 form to create knockouts in the hybrid poplar clone“717-1B4”(*** x *** clone INRA 717-1B4).Alternative CRISPR/Cas9-based technologies,*** involving modified Cas9 for gene activation and base editing,have not been evaluated in most Populus species for their *** we employed a deactivated Cas9(dCas9)-based CRISPR activation(CRISPRa)technique to fine-tune the expression of two target genes,TPX2 and LecRLK-G which play important roles in plant growth and defense response,in hybrid poplar clone“717-1B4”and poplar clone“WV94”(***“WV94”),*** observed that CRISPRa resulted in 1.2-fold to 7.0-fold increase in target gene expression through transient expression in protoplasts and Agrobacterium-mediated stable transformation,demonstrating the effectiveness of dCas9-based CRISPRa system in *** addition,we applied Cas9 nickase(nCas9)-based cytosine base editor(CBE)to precisely introduce premature stop codons via C-to-T conversion,with an efficiency of 13%–14%,in the target gene PLATZ which encodes a transcription factor involved in plant fungal pathogen response in hybrid poplar clone“717-1B4”.Overall,we showcase the successful application of CRISPR/Cas-based technologies in gene expression regulation and precise gene engineering in two Populus species,facilitating the adoption of emerging genome editing tools in woody species.

关键词： CRISPR/Cas9 activation base

Different conformational responses of the β2-adrenergic receptor-Gs complex upon binding of the partial agonist salbutamol or the full agonist isoprenaline

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National Science Review 2021年第9期8卷 111-120页

作者： Fan Yang Shenglong Ling Yingxin Zhou Yanan zhang Pei Lv Sanling Liu Wei fang Wenjing Sun Liaoyuan A.Hu Longhua zhang Pan Shi Changlin Tian Hefei National Laboratory of Physical Sciences at Microscale and School of Life Sciences University of Science and Technology of China Amgen Asia R&D Center Amgen Research Shanghai 201210 China3High Magnetic Field Laboratory Chinese Academy of Sciences

G protein-coupled receptors(GPCRs) are responsible for most cytoplasmic signaling in response to extracellular ligands with different efficacy profiles. Various spectroscopic techniques have identified that agonists exhibiting varying efficacies can selectively stabilize a specific conformation of the ***, the structural basis for activation of the GPCR-G protein complex by ligands with different efficacies is incompletely understood. To better understand the structural basis underlying the mechanisms by which ligands with varying efficacies differentially regulate the conformations of receptors and G proteins, we determined the structures of β2AR-Gαsβγ bound with partial agonist salbutamol or bound with full agonist isoprenaline using single-particle cryo-electron microscopy at resolutions of 3.26 ?A and3.80 ?A, respectively. Structural comparisons between the β2AR-Gs-salbutamol and β2AR-Gs-isoprenaline complexes demonstrated that the decreased binding affinity and efficacy of salbutamol compared with those of isoprenaline might be attributed to weakened hydrogen bonding interactions, attenuated hydrophobic interactions in the orthosteric binding pocket and different conformational changes in the rotamer toggle switch in TM6. Moreover, the observed stronger interactions between the intracellular loop 2 or 3(ICL2 or ICL3) of β2AR and Gαswith binding of salbutamol versus isoprenaline might decrease phosphorylation in the salbutamol-activated β2AR-Gs complex. From the observed structural differences between these complexes of β2AR, a mechanism of β2AR activation by partial and full agonists is proposed to provide structural insights into β2AR desensitization.

关键词： cryo-EM structure G protein-coupled receptor (GPCR) partial and full agonists conformational change desensitization

Dietary Lipid Intervention in the Prevention of Brain Aging

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Engineering 2024年第6期37卷 128-137页

作者： Wei Xiong Bing fang Xiaoyu Wang Ming zhang Min Du Jiazeng Sun Juan Chen Yixuan Li Changhao Sun Xingen Lei Xue zhang Fazheng Ren Key Laboratory of Precision Nutrition and Food Quality Department of Nutrition and HealthChina Agricultural UniversityBeijing 100083China Key Laboratory of Functional Dairy Co-Constructed by the Ministry of Education and Beijing MunicipalityCollege of Food Science and Nutritional EngineeringChina Agricultural UniversityBeijing 100083China School of Food Science and Chemical Engineering Beijing Technology and Business UniversityBeijing 100048China Department of Animal Science Washington State UniversityPullmanWA 99164USA Department of Nutrition and Food Hygiene School of Public HealthHarbin Medical UniversityHarbin 150028China Department of Animal Science Cornell UniversityIthacaNY 14853USA National Health Commission(NHC)and Chinese Academy of Medical Sciences(CAMS)Key Laboratory of Molecular Probe and Targeted Theranostics Harbin Medical UniversityHarbin 150028China

As people live longer,the burden of aging-related brain diseases,especially dementia,is *** aging increases the risk of cognitive impairment,which manifests as a progressive loss of neuron function caused by the impairment of synaptic plasticity via disrupting lipid ***,supplemental dietary lipids have the potential to prevent brain *** review summarizes the important roles of dietary lipids in brain function from both structure and mechanism *** and animal studies have provided evidence of the functions of polyunsaturated fatty acids(PUFAs)in brain *** results of interventions indicate that phospholipids—including phosphatidylcholine,phosphatidylserine,and plasmalogen—are efficient in alleviating cognitive impairment during aging,with plasmalogen exhibiting higher efficacy than *** is a recognized nutrient used in clinical trials due to its special vinyl ether bonds and abundance in the postsynaptic membrane of *** research should determine the dose-dependent effects of plasmalogen in alleviating brain-aging diseases and should develop extraction and storage procedures for its clinical application.

关键词： Brain aging Nutritional intervention Phospholipids Phosphatidylcholine Phosphatidylserine Plasmalogen

High Quantum Efficiency Back-Illuminated AlGaN-Based Solar-Blind Ultraviolet p-i-n Photodetectors

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Chinese Physics Letters 2012年第9期29卷 166-169页

作者： WANG Guo-Sheng LU Hai XIE Feng CHEN Dun-Jun REN fang-fang zhang Rong ZHENG You-Dou Jiangsu Provincial Key Laboratory of Advanced Photonic and Electronic Materials and School of Electronic Science and EngineeringNanjing UniversityNanjing 210093

AlGaN-based back-illuminated solar-blind ultraviolet(UV)p–i–n photodetectors(PDs)with high quantum efficiency are fabricated on sapphire *** improve the overall performance of the PD,a series of structural design considerations and growth procedures are implemented in the epitaxy process.A distinct wavelength-selective photo-response peak of the PD is obtained in the solar-blind *** operating in photovoltaic mode,the PD exhibits a solar-blind/UV rejection ratio of up to 4 orders of magnitude and a peak responsivity of~113.5 mA/W at 270 nm,which corresponds to an external quantum efficiency of~52%.Under a reverse bias of-5 V,the PD shows a low dark current of~1.8 pA and an enhanced peak quantum efficiency of~64%.The thermal noise limited detectivity is estimated to be~3.3×10^(13) cm·Hz^(1/2)W^(-1).

关键词： AlGaN Efficiency sapphire

High-pressure synergetic measurement station(HP-SymS)

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Chinese Physics B 2018年第7期27卷 8-19页

作者： Xiaohui Yu fangfei Li Yonghao Han fang Hong Changqing jin Zhi He Qiang Zhou Beijing National Laboratory for Condensed Matter Physics Institute of PhysicsChinese Academy of SciencesBeijing 100190China State Key Laboratory of Superhard Materials College of PhysicsJilin UniversityChangchun 130012China

In the High-Pressure Synergetic Measurements Station （HP-SymS） of the Synergic Extreme Condition User Facility （SECUF）, we will develop ultrahigh-pressure devices based on diamond-anvil cell （DAC） techniques, with a target pressure up to 300 GPa. With the use of cryostat and magnet, we will reach 300 GPa4.2 K-9 T and conduct simultaneous measurements of the electrical-transport property and Raman/Brillouin spectrascopy. With resistance heating and laser heating, we will reach temperatures of at least 1000 and 3000 K, respectively, coupled with Raman/Brillouin spectroscopy measurements. Some designs of supporting devices, such as a femtosecond laser gasket-drilling device, electrode-deposition device, and the gas-loading device, are also introduced in this article. Finally, we conclude by providing some perspectives on the applications of the DAC in related research fields.

关键词： high pressure synergetic measurements diamond-anvil cell (DAC)

East Asian monsoon intensification promoted weathering of the magnesium-rich southern China upper crust and its global significance

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Science China Earth Sciences 2021年第7期64卷 1155-1170页

作者： Yibo YANG Albert GALY Xiaomin fang Christian FRANCE-LANORD Shiming WAN Rongsheng YANG Jian zhang Ran zhang Song YANG Yunfa MIAO Yudong LIU Chengcheng YE Key Laboratory of Continental Collision and Plateau Uplift Institute of Tibetan Plateau ResearchChinese Academy of SciencesBeijing 100101China CAS Center for Excellence in Tibetan Plateau Earth Sciences Beijing 100101China Centre de Recherches Pétrographiques et Géochimiques UMR7358CNRS-Universitéde LorraineVandoeuvre les Nancy 54500France University of Chinese Academy of Sciences Beijing 100049China Key Laboratory of Marine Geology and Environment Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory for Marine Geology Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237China Climate Change Research Center Institute of Atmospheric PhysicsChinese Academy of SciencesBeijing 100029China Key Laboratory of Tibetan Environment Changes and Land Surface Processes Institute of Tibetan Plateau ResearchChinese Academy of SciencesBeijing 100101China Key Laboratory of Desert and Desertification Northwest Institute of Eco-Environment and ResourcesChinese Academy of SciencesLanzhou 730000China

The Oligocene-Miocene boundary Asian climatic reorganization linked to the northward migration of the East Asian monsoon into subtropical China is a potentially important but poorly constrained atmospheric CO_(2) consumption ***,we performed a first-order estimate of the CO_(2) consumption induced by silicate chemical weathering and organic carbon burial in subtropical China related to this climatic *** results show that an increase in long-term CO_(2 )consumption by silicate weathering varies from 0.06×10^(12)to 0.87×10^(12)mol yr^(-1)depending on erosion flux reconstructions,with an~50%contribution of Mg-silicate weathering since the late *** organic carbon burial flux is approximately 25%of the contemporary CO_(2) consumption by silicate *** results highlight the significant role of weathering of the Mg-rich upper continental crust in East China,which would contribute to the rapid decline in atmospheric CO_(2) during the late Oligocene and the Neogene rise in the seawater Mg *** this climatic reorganization was mainly induced by the Tibetan Plateau uplift,our study suggests that the growth of the Himalayan-Tibetan Plateau can lead to indirect modification of the global carbon and magnesium cycles by changing the regional hydrological cycle in areas of East Asia that are tectonically less active.

关键词： Tibetan Plateau Late Oligocene Silicate weathering Carbon cycle Oceanic Mg cycle

Synthesis and fluorescence properties of CdTe:Eu3+ nanocrystals and core-shell SiO2-coated CdTe:Eu3+ nanospheres

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Rare Metals 2019年第10期38卷 989-995页

作者： Jun-fang Gao Jun-Hong Yang Xiao-Yong zhang Jun Zhao Xing-Wang Liu Bing-Feng Shi Key Laboratory of Materials Physical Chemistry Baotou Teachers CollegeBaotou 014000China Institute of Chemistry Northeast Normal UniversityChangchun 130000China

Eu3+ doped-CdTe(CdTe:Eu3+)nanocrystals were prepared via a facile hydrothermal method,and Eu3+ was successfully incorporated into the crystal lattice of CdTe and measured by X-ray powder diffraction(XRD),transmission electron microscopy(TEM),ultraviolet-visible(UV-Vis) absorption spectroscopy and fluorescence *** CdTe:Eu^3+ nanocrystals still have a cubic crystal structure,and the corresponding XRD peaks of CdTe:Eu3+nanocrystals shift to larger angles compared with those of pure *** CdTe:Eu3+ nanocrystals are monodisperse and the particles size is about 2-4 *** with pure CdTe,the CdTe:Eu^3+ nanocrystals have larger band gap and thus exhibit blueshift in the emission spectra,which could be accounted for by the energy transfer between Eu^3+ and *** enhance the stability and functionality of CdTe:Eu3+nanocrystals,the CdTe:Eu3+nanocrystals were coated with SiO2 and the core-shell SiO2-coated CdTe:Eu3+nanocrystals(CdTe:Eu^3+@SiO2) were prepared via microemulsion *** results show that CdTe:Eu3+nanocrystals are uniformly dispersed in the shell,and CdTe:Eu3+@SiO2 nanospheres are uniformly spherical with an average diameter of about 75 *** fluorescence emission of CdTe:Eu3+@SiO2(567 nm) shows a blueshift compared with that of CdTe:Eu^3+nanocrystals(632 nm),possibly because of altered surface properties after SiO2 ***:Eu^3+and CdTe:Eu^3+@SiO2 with tunable photoluminescence are potentially useful in fabricating optical and bioimaging devices.

关键词： CdTe:Eu^3+nanocrystals Doping Core-shell SiO2 Fluorescent properties

SCAG:A Stratified,Clustered,and Growing-Based Algorithm for Soybean Branch Angle Extraction and Ideal Plant Architecture Evaluation

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Plant Phenomics 2024年第3期6卷 595-609页

作者： Songyin zhang Yinmeng Song Ran Ou Yiqiang Liu Shaochen Li Xinlan Lu Shan Xu Yanjun Su Dong Jiang Yanfeng Ding Haifeng Xia Qinghua Guo jin Wu Jiaoping zhang Jiao Wang Shichao jin Plant Phenomics Research Centre Academy for Advanced Interdisciplinary StudiesCollaborative Innovation Centre for Modern Crop Production cosponsored by Province and MinistryState Key Laboratory of Crop Genetics and Germplasm EnhancementNanjing Agricultural UniversityNanjing 210095China National Center for Soybean Improvement Key Laboratory for Biology and Genetic Improvement of Soybean(GeneralMinistry of Agriculture)College of AgricultureNanjing Agricultural UniversityNanjing 210095China Sanya Research Institute of Nanjing Agriculture University Sanya 572024China State Key Laboratory of Vegetation and Environmental Change Institute of BotanyChinese Academy of SciencesBeijing 100093China School of Automation Southeast UniversityNanjing 210096China Institute of Remote Sensing and Geographic Information System School of Earth and Space SciencesPeking UniversityBeijing 100871China Division for Ecology and Biodiversity School of Biological SciencesThe University of Hong KongHong KongChina

Three-dimensional(3D)phenotyping is important for studying plant structure and *** detection and ranging(LiDAR)has gained prominence in 3D plant phenotyping due to its ability to collect 3D point ***,organ-level branch detection remains challenging due to small targets,sparse points,and low signal-to-noise *** addition,extracting biologically relevant angle traits is *** this study,we developed a stratified,clustered,and growing-based algorithm(SCAG)for soybean branch detection and branch angle calculation from LiDAR data,which is heuristic,open-source,and *** achieved high branch detection accuracy(F-score=0.77)and branch angle calculation accuracy(r=0.84)when evaluated on 152 diverse soybean ***,the SCAG outperformed 2 other classic algorithms,the support vector machine(F-score=0.53)and density-based methods(F-score=0.55).Moreover,after applying the SCAG to 405 soybean varieties over 2 consecutive years,we quantified various 3D traits,including canopy width,height,stem length,and average *** data filtering,we identified novel heritable and repeatable traits for evaluating soybean density tolerance potential,such as the ratio of average angle to height and the ratio of average angle to stem length,which showed greater potential than the well-known ratio of canopy width to height *** work demonstrates remarkable advances in 3D phenotyping and plant architecture *** algorithm can be applied to other crops,such as maize and *** dataset,scripts,and software are public,which can further benefit the plant science community by enhancing plant architecture characterization and ideal variety selection.

关键词： architecture evaluation algorithm angle plant branch ideal clustered extraction growing-based

Radix Kansui Stir-Fried with Vinegar Reduces Radix Kansui-Related Hepatotoxicity in Mice via Mitochondrial Pathway

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Chinese Journal of Integrative Medicine 2021年第3期27卷 192-197页

作者： BAO Bei-hua YAN Xiao-jing CAO Yu-dan YAO Wei-feng CHENG fang-fang CHEN Pei-dong SHAN Ming-qiu zhang Li DING An-wei Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative MedicineNanjing University of Chinese MedicineNanjing 210023China Menghe Medical Institute Changzhou Traditional Chinese Medical Hospital Affiliated to Nanjing University of Chinese MedicineChangzhouJiangsu Province 213003China

Objective To investigate the mechanism of Radix Kansui(RK)stir-fried with vinegar(VRK)decreased hepatotoxicity in *** According to a random number table,40 mice were randomly divided into negative control group(0.5%carboxymethylcellulose sodium,20 mL/kg),positive control group(0.1%mixture of carbon tetrachloride in soybean oil,20 mL/kg),RK group(the ethyl acetate extracts of RK,250 g crude drug/kg)and VRK group(the ethyl acetate extracts of VRK,250 g crude drug/kg)with 10 mice per *** mice were administered orally by gavage daily for 7 continuous *** morphology of liver tissues was examined to assess the liver injury by a transmission electron *** apoptosis in vivo was determined by terminal deoxynucleotidyltransferase-mediated dUTP-biotin nickend labeling(TUNEL)*** technique was adopted to detect the expression of particular antiapoptotic and proapoptotic proteins in the mitochondrial pathways,including B-cell lymphoma(Bcl-2)and caspase-3,as well as the expression of inflammatory mediators,including nuclear factor kappa B(NF-κB)and intercellular adhesion molecule-1(ICAM-1).Results Liver injury and hepatocyte apoptosis were observed in RK mice,and the liver injury were significantly reduced in VRK-treated *** immunohistochemistry study,compared with the negative control group,RK inhibited dramatically the Bcl-2 protein expression and significantly increased the expression of caspase-3,NF-κB and ICAM-1(all P<0.01).Compared with the RK group,VRK group induced significant increase on Bcl-2 protein expression,and decreased the caspase-3,NF-κB and ICAM-1 protein expression(P<0.05 or <0.01).Conclusion The mechanism of reduced hepatotoxicity of VRK may be associated with the reduced inflammation,regulation of antiapoptotic and proapoptotic mediators in the mitochondrial pathway.

关键词： Radix Kansui mitochondrial pathway hepatotoxicity apoptosis inflammation Chinese medicine