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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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“Three medicines and three formulas”in COVID-19:from bench to bedside 被引量:1
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作者 Guangxu Xiao Simeng Wang +6 位作者 Ruijiao Wang Li Tan Shibo Ma Shuang He Guanwei Fan Yan Zhu ming lyu 《Acupuncture and Herbal Medicine》 2023年第4期309-322,共14页
Chinese guideline has proven effective in the fight against coronavirus disease 2019(COVID-19)during the global spread of the epidemic.Traditional Chinese medicine(TCM)is widely recognized for its effectiveness in all... Chinese guideline has proven effective in the fight against coronavirus disease 2019(COVID-19)during the global spread of the epidemic.Traditional Chinese medicine(TCM)is widely recognized for its effectiveness in alleviating symptoms,inhibiting disease progress,reducing mortality,and improving the cure rate of patients with COVID-19.During the pandemic,“three medicines and three formulas”(TMTFs)stood out from hundreds of others in registered clinical studies,which became highly recommended as TCM for COVID-19 treatment.The TMTFs not only effectively relieve the clinical symptoms of fever,cough,fatigue,and phlegm,but also substantially shorten the time of nucleic acid negative conversion,improve lung computed tomography imaging features and inflammation,ameliorate clinical biochemical indicators,and reduce sequelae.Their potential pharmacological mechanisms are mainly related to the crosstalk between viral toxicity,endothelial damage,cytokine storm,immune response,and microthrombus.In brief,the clinical effects and the potential mechanisms of TMTFs on COVID-19 were systematically analyzed and summarized covering the entirety of disease development,including virus invasion and replication,immune response and cytokine storm,and acute respiratory distress syndrome and multiple organ dysfunction syndrome.This review provides a theoretical basis and reference for the in-depth understanding of the positive role of TMTFs in COVID-19 treatment. 展开更多
关键词 Clinical research COVID-19 Potential mechanism SARS-CoV-2 Three medicines and three formulas
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Traditional Chinese medicine in COVID-19 被引量:25
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作者 ming lyu Guanwei Fan +12 位作者 Guangxu Xiao Taiyi Wang Dong Xu Jie Gao Shaoqin Ge Qingling Li Yuling Ma Han Zhang Jigang Wang Yuanlu Cui Junhua Zhang Yan Zhu Boli Zhang 《Acta Pharmaceutica Sinica B》 SCIE CAS CSCD 2021年第11期3337-3363,共27页
COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2(SARSCo V-2)has spread across the globe,posing an enormous threat to public health and safety.Traditional Chinese medicine(TCM),in combination... COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus 2(SARSCo V-2)has spread across the globe,posing an enormous threat to public health and safety.Traditional Chinese medicine(TCM),in combination with Western medicine(WM),has made important and lasting contributions in the battle against COVID-19.In this review,updated clinical effects and potential mechanisms of TCM,presented in newly recognized three distinct phases of the disease,are summarized and discussed.By integrating the available clinical and preclinical evidence,the efficacies and underlying mechanisms of TCM on COVID-19,including the highly recommended three Chinese patent medicines and three Chinese medicine formulas,are described in a panorama.We hope that this comprehensive review not only provides a reference for health care professionals and the public to recognize the significant contributions of TCM for COVID-19,but also serves as an evidence-based in-depth summary and analysis to facilitate understanding the true scientific value of TCM. 展开更多
关键词 COVID-19 SARS-CoV-2 Traditional Chinese medicine Clinical evidence Potential mechanism Viral infection Cytokine storm Multiple organ dysfunction
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