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不同CRISPR/Cas9供体适配基因编辑系统的比较及优化研究
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作者 马宝霞 杨森 +6 位作者 吕明 王昱人 常立业 韩艺帆 王建刚 郭杨 徐坤 《遗传》 CAS CSCD 北大核心 2024年第6期466-477,共12页
在哺乳动物细胞中进行基因敲入通常采用同源定向修复(homology-directed repair,HDR)机制将外源DNA模板整合到目标基因组靶点中。然而HDR效率往往较低,其中外源DNA模板与目标基因组靶点的共定位是关键限制因素之一。为提高CRISPR/Cas9... 在哺乳动物细胞中进行基因敲入通常采用同源定向修复(homology-directed repair,HDR)机制将外源DNA模板整合到目标基因组靶点中。然而HDR效率往往较低,其中外源DNA模板与目标基因组靶点的共定位是关键限制因素之一。为提高CRISPR/Cas9系统介导的HDR效率,本团队及前人研究将不同接头蛋白与SpCas9蛋白融合表达,利用其与特异性DNA序列结合的特性,构建了多种CRISPR/SpCas9供体适配基因编辑系统。为了便于比较、优化不同CRISPR/Cas9供体适配系统,本研究利用这些系统在HEK293T细胞GAPDH和ACTB基因末位外显子3′-端进行了eGFP基因敲入,并采用了优化的供体DNA模板设计方式,通过PCR和Sanger测序检测敲入的准确性,流式细胞分析进行敲入效率的检测。结果表明,将yGal4BD、hGal4BD、hLacI、hTHAP11和N57等接头蛋白与SpCas9蛋白C-端融合对其活性均无显著性影响;在GAPDH位点上,SpCas9融合yGal4BD、hGal4BD、hLacI和hTHAP11的供体适配系统等均能显著提高敲入效率;在ACTB位点上,SpCas9融合yGal4BD和hGal4BD能显著提高敲入效率;且增加供体DNA模板中的结合序列(binding sequence,BS)数量,有利于提高SpCas9-hTHAP11系统介导的敲入效率。总之,本研究比较并优化了不同的CRISPR/Cas9供体适配基因编辑系统,为后续相关的基因编辑应用研究提供了参考和借鉴。 展开更多
关键词 基因编辑 基因敲入 CRISPR/Cas9 供体适配 同源定向修复
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Preparation of Cu-based Bulk Metallic Glass Matrix Composites 被引量:1
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作者 Yufeng SUN yuren wang +1 位作者 Bingchen WEI Weihuo LI 《Journal of Materials Science & Technology》 SCIE EI CAS CSCD 2006年第1期73-77,共5页
Cu47Ti34Zr11Ni8 bulk metallic glass (BMG) matrix composites containing in situ formed TiC particles and 5-TiCu dendrite phase were developed by copper mold cast. The thermal stability and microstructure of the compo... Cu47Ti34Zr11Ni8 bulk metallic glass (BMG) matrix composites containing in situ formed TiC particles and 5-TiCu dendrite phase were developed by copper mold cast. The thermal stability and microstructure of the composites are investigated. Room temperature compression tests reveal that the composite samples exhibit higher fracture strength and distinct plastic strain of 0.2%-0.5%, comparing with that of the corresponding Cu47Ti34Zr11Ni8 monolithic BMG. 展开更多
关键词 Bulk metallic glasses Shear bands Mechanical properties COMPOSITES
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PSA controls hepatic lipid metabolism by regulating the NRF2 signaling pathway 被引量:4
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作者 Bangliang Huang Xin Xiong +8 位作者 Linlin Zhang Xiufei Liu yuren wang Xiaoli Gong Qian Sang Yongling Lu Hua Qu Hongting Zheng Yi Zheng 《Journal of Molecular Cell Biology》 SCIE CAS CSCD 2021年第7期527-539,共13页
The activity of proteinase is reported to correlate with the development and progression of nonalcoholic fatty liver disease(NAFLD).Puromycin-sensitive aminopeptidase(PSA/NPEPPS)is an integral nontransmembrane enzyme ... The activity of proteinase is reported to correlate with the development and progression of nonalcoholic fatty liver disease(NAFLD).Puromycin-sensitive aminopeptidase(PSA/NPEPPS)is an integral nontransmembrane enzyme that functions to catalyze the cleavage of amino acids near the N-terminus of polypeptides.A previous study suggested that this enzyme acts as a regulator of neuropeptide activity;however,the metabolic function of this enzyme in the liver has not been explored.Here,we identified the novel role of PSA in hepatic lipid metabolism.Specifically,PSA expression was lower in fatty livers from NAFLD patients and mice(HFD,ob/ob,and db/db).PSA knockdown in cultured hepatocytes exacerbated diet-induced triglyceride accumulation through enhanced lipogenesis and attenuated fatty acid β-oxidation.Moreover,PSA mediated activation of the master regulator of antioxidant response,nuclear factor erythroid 2-related factor 2(NRF2),by stabilizing NRF2 protein expression,which further induced downstream antioxidant enzymes to protect the liver from oxidative stress and lipid overload.Accordingly,liver-specific PSA overexpression attenuated hepatic lipid accumulation and steatosis in ob/ob mice.Furthermore,in human liver tissue samples,decreased PSA expression correlated with the progression of NAFLD.Overall,our findings suggest that PSA is a pivotal regulator of hepatic lipid metabolism and its antioxidant function occurs by suppressing NRF2 ubiquitination.Moreover,PSA may be a potential biomarker and therapeutic target for treating NAFLD. 展开更多
关键词 PSA NAFLD LIPOGENESIS fatty acidβ-oxidation NRF2
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Microrheological Insights into the Dynamics of Amyloplasts in Root Gravity-Sensing Cells 被引量:1
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作者 Zhongyu Zheng Junjie Zou +3 位作者 Hanhai Li Shan Xue yuren wang Jie Le 《Molecular Plant》 SCIE CAS CSCD 2015年第4期660-663,共4页
Gravitropism in plants is key for orienting organs such as the Darwin's description of the opposite growth direction of roots and shoots in his book The Power of Movement in Plants more than 100 years ago (Darwin, 1... Gravitropism in plants is key for orienting organs such as the Darwin's description of the opposite growth direction of roots and shoots in his book The Power of Movement in Plants more than 100 years ago (Darwin, 1880). The gravitropic response of an oriented plant is divided into three sequential phases: gravity sensing, signal transmission, and the growth response. As the initial phase of gravitropism, gravity sensing (the conversion of the mechanical stimulus into a biochemical signal) has been intensively studied, but much is still unknown. The statoliths are vital for sense of balance and response to gravity, e.g. in statocytes for invertebrates and in inner ears for vertebrates. 展开更多
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High-end equipment customer requirement analysis based on opinion extraction
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作者 Yuejin TAN yuren wang +2 位作者 Xin LU Mengsi CAI Bingfeng GE 《Frontiers of Engineering Management》 2018年第4期479-486,共8页
Acquisition and analysis of customer requirements are the essential steps in high-end equipment design.Considering that Internet and big data technologies are integrated into the manufacturing industry, we propose a m... Acquisition and analysis of customer requirements are the essential steps in high-end equipment design.Considering that Internet and big data technologies are integrated into the manufacturing industry, we propose a method of analyzing customer requirements based on open-source data. First, online data are collected with focused crawlers and preprocessed to filter noise and duplicate. Then, user opinions are extracted based on the defined template, and users' sentiments are analyzed.Based on the relationship between user sentiments and attribute parameters, the parameter range that satisfies customers can be obtained. The proposed method is evaluated by using an example of new energy vehicle to verify its availability and feasibility. 展开更多
关键词 REQUIREMENT analysis OPINION EXTRACTION HIGH-END EQUIPMENT new energy vehicle
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Structural studies on the disorder system of liquid gallium
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作者 Shiqiang Wei Wenhan Liu +2 位作者 Tiandou Hu yuren wang Kunquan Lu 《Chinese Science Bulletin》 SCIE EI CAS 1998年第5期396-398,共3页
关键词 LIQUID GA XAFS DISORDER system.
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