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Base editors:development and applications in biomedicine

Frontiers of Medicine 2023年第3期17卷 359-387页

作者： Yanhui Liang Fangbing Chen kepin wang Liangxue Lai China-New Zealand Joint Laboratory on Biomedicine and Health CAS Key Laboratory of Regenerative BiologyGuangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineCentre for Regenerative Medicine and HealthHong Kong Institute of Science and InnovationGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China Research Unit of Generation of Large Animal Disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou 510530China Sanya Institute of Swine Resource Hainan Provincial Research Centre of Laboratory AnimalsSanya 572000China Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine Wuyi UniversityJiangmen 529020China

Base editor(BE)is a gene-editing tool developed by combining the CRISPR/Cas system with an individual deaminase,enabling precise single-base substitution in DNA or RNA without generating a DNA double-strand break(DSB)or requiring donor DNA templates in living *** editors offer more precise and secure genome-editing effects than other conventional artificial nuclease systems,such as CRISPR/Cas9,as the DSB induced by Cas9 will cause severe damage to the ***,base editors have important applications in the field of biomedicine,including gene function investigation,directed protein evolution,genetic lineage tracing,disease modeling,and gene *** the development of the two main base editors,cytosine base editors(CBEs)and adenine base editors(ABEs),scientists have developed more than 100 optimized base editors with improved editing efficiency,precision,specificity,targeting scope,and capacity to be delivered in vivo,greatly enhancing their application potential in ***,we review the recent development of base editors,summarize their applications in the biomedical field,and discuss future perspectives and challenges for therapeutic applications.

关键词： base editing CBE ABE ADAR DdCBE disease model therapeutic application

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In vivo evaluation of guide-free Cas9-induced safety risks in a pig model

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Signal Transduction and Targeted Therapy 2024年第8期9卷 3493-3506页

作者： Weikai Ge Shixue Gou Xiaozhu Zhao Qin Jin Zhenpeng Zhuang Yu Zhao Yanhui Liang Zhen Ouyang Xiaoyi Liul Fangbing Chen Hui Shi Haizhao Yan Han Wu Liangxue Lai kepin wang China-New Zealand Joint Laboratory on Biomedicine and Health CAS Key Laboratory of Regenerative BiologyGuangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineJoint School of Life SciencesGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou Medical UniversityGuangzhou 510530China Sanya institute of Swine resource Hainan Provincial Research Center of Laboratory AnimalsSanya 572000China Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine Wuyi UniversityJiangmen 529020China Guangzhou National Laboratory Guangzhou 510005China SUniversity of Chinese Academy of Sciences 100049 BeijingChina Research Unit of Generation of Large Animal Disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou 510530China

The CRISPR/Cas9 system has shown great potential for treating human genetic diseases through gene ***,there are concerns about the safety of this system,specifically related to the use of guide-free *** studies have shown that guidefree Cas9 can induce genomic instability in ***,the in vivo safety risks associated with guide-free Cas9 have not been evaluated,which is necessary for the development of gene therapy in clinical *** this study,we used doxycycline-inducible Cas9-expressing pigs to evaluate the safety risks of guide-free Cas9 in *** findings demonstrated that expression of guide-free Cas9 could induce genomic damages and transcriptome changes in *** severity of the genomic damages and transcriptome changes were correlate with the expression levels of Cas9 ***,prolonged expression of Cas9 in pigs led to abnormal phenotypes,including a significant decrease in body weight,which may be attributable to genomic damage-induced nutritional absorption and metabolic ***,we observed an increase in whole-genome and tumor driver gene mutations in pigs with long-term Cas9 expression,raising the risk of tumor *** in vivo evaluation of guide-free Cas9 in pigs highlights the necessity of considering and monitoring the detrimental effects of Cas9 alone as genome editing via the CRISPR/Cas9 system is implemented in clinical gene *** research emphasizes the importance of further study and implementation of safety measures to ensure the successful and safe application of the CRiSPR/Cas9 system in clinical practice.

关键词： CRISPR/Cas9 vivo alone

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Improving the Cpf1-mediated base editing system by combining dCas9/dead sgRNA with human APOBEC3A variants

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Journal of Genetics and Genomics 2021年第1期48卷 92-95页

作者： Meng Lian Fangbing Chen Xingyun Huang Xiaozhu Zhao Shixue Gou Nan Li Qin Jin Hui Shi Yanhui Liang Jingke Xie Weikai Ge Zhenpeng Zhuang Jiaowei wang Yinghua Ye Yi Yang kepin wang Liangxue Lai Han Wu CAS Key Laboratory of Regenerative Biology Guangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China Institute of Physical Science and Information Technology Anhui UniversityHefei 230601China University of Chinese Academy of Sciences Beijing 100049China Research Unit of Generation of Large Animal Disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou 510530China Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory) Guangzhou 510005China Key Laboratory for Major Obstetrics Diseases of Guangdong Province Key Laboratory of Reproduction and Genetics of Guangdong Higher Education InstitutesThe Third Hospital Affiliated of Guangzhou Medical UniversityGuangzhou 510150China

Base editor-mediated C-to-T base conversion obviates the requirements of double-strand breaks,thereby showing promise as a tool for disease modeling and gene therapy(Gaudelli et al.,2017;Rees and Liu 2018).The most actively used base editor comprises a Cas9 nickase(nCas9)with cytidine deaminase and fused uracil DNA glycosylase inhibitor at the carboxy terminus of nCas9 to inhibit uracil N-glycosylase effects(Pearl,2000;Kunz et al.,2009;Rees and Liu 2018).

关键词： base uracil thereby

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Genetically humanized pigs exclusively expressing human insulin are generated through custom endonuclease-mediated seamless engineering

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Journal of Molecular Cell Biology 2016年第2期8卷 174-177页

作者： Yi Yang kepin wang Han Wu Qin Jin Degong Ruan Zhen Ouyang Bentian Zhao Zhaoming Liu Yu Zhao Quanjun Zhang Nana Fan Qishuai Liu Shimei Guo Lei Bu Yong Fan Xiaofang Sun Xiaoping Li Liangxue Lai Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine South China Institute for Stem Cell Biology and Regenerative MedicineGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China School of Life Sciences University of Science and Technology of ChinaHefei 230027China Leon H Charney Division of Cardiology New York University School of Medicine522 First AvenueNew YorkNY 10016USA Key Laboratory for Major Obstetric Diseases of Guangdong Province Key Laboratory of Reproduction and Genetics of Guangdong Higher Education InstitutesThe Third Affiliated Hospital of Guangzhou Medical UniversityGuangzhou 510150China Jilin Provincial Key Laboratory of Animal Embryo Engineering Institute of ZoonosisCollege of Veterinary MedicineJilin UniversityChangchun 130062China

Dear Editor,Type1diabetes(T1D)isa lifelong(chronic)disease and a major health problem throughout the *** disease can be treatedby either insulin injection or islet *** transplantation is considered as a better treatment for T1D patients,because islets can produce and release insulin at the appropriate time,resulting in tight blood glucose control.

关键词： islet injection expressing

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Double knock-in pig models with elements of binary Tet-On and phiC31 integrase systems for controllable and switchable gene expression

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Science China(Life Sciences) 2022年第11期65卷 2269-2286页

作者： Qin Jin Xiaoyu Yang Shixue Gou Xiaoyi Liu Zhenpeng Zhuang Yanhui Liang Hui Shi Jiayuan Huang Han Wu Yu Zhao Zhen Ouyang Quanjun Zhang Zhaoming Liu Fangbing Chen Weikai Ge Jingke Xie Nan Li Chengdan Lai Xiaozhu Zhao Jiaowei wang Meng Lian Lei Li Longquan Quan Yinghua Ye Liangxue Lai kepin wang CAS Key Laboratory of Regenerative Biology Joint School of Life SciencesGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou Medical UniversityGuangzhou 510530China Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine Centre for Regenerative Medicine and HealthHong Kong Institute of Science and InnovationGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou 510530China Sanya institute of Swine resource Guangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesHainan Provincial Research Center of Laboratory AnimalsSanya 572000China Research Unit of Generation of Large Animal Disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou 510530China University of Chinese Academy of Sciences Beijing 100049China Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory) Guangzhou 510005China Guangdong Provincial Key Laboratory of Laboratory Animals Guangdong Laboratory Animals Monitoring InstituteGuangzhou 510633China Guangdong Provincial Key Laboratory of Large Animal models for Biomedicine Wuyi UniversityJiangmen 529020China

Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological *** Tet-On system has been widely employed to regulate transgenic expression by *** pig models with tetracycline regulatory elements were generated through random *** process often resulted in uncertain expression and unpredictable phenotypes,thus hindering their ***,by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus,respectively,a double knock-in reporter pig model was *** characterized excellent properties of this system for controllable transgenic expression both in vitro and in *** att P sites were arranged to flank the td Tomato to switch reporter *** or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer *** display the flexible application of this system,we generated a pig strain with Dox-inducing h KRASexpression through phiC31 integrase-mediated cassette *** eight months of Dox administration,squamous cell carcinoma developed in the nose,mouth,and scrotum,which indicated this pig strain could serve as an ideal large animal model to study ***,the established pig models with controllable and switchable transgene expression system will provide a facilitating platform for transgenic and biomedical research.

关键词： double knock-in Tet-On system phiC31-mediated cassette exchange pig model hKRASG12D-related tumorigenesis

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基于“互联网+”的虚拟现实(VR)计算机辅助教学系统设计

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科教导刊 2019年第18期 129-130页

作者：刘泽宇张思维陆觐万航王可拼王亚莉江苏第二师范学院现代传媒学院

目前主流的课件制作方式主要是幻灯片或者动画,对于图文信息呈现时直观度有限。虚拟现实技术作为创建模拟环境的计算机系统,能够给用户身临其境的沉浸式体验。利用虚拟现实技术设计虚拟现实计算机辅助教学系统,教师使用系统设计制作课件... 详细信息

目前主流的课件制作方式主要是幻灯片或者动画,对于图文信息呈现时直观度有限。虚拟现实技术作为创建模拟环境的计算机系统,能够给用户身临其境的沉浸式体验。利用虚拟现实技术设计虚拟现实计算机辅助教学系统,教师使用系统设计制作课件,有效实现将学习内容以可感的方式呈现给学习者。其分为教师端和学生端。教师端主要功能为VR课件的制作和上传。学生端能够实现VR课件播放,教师端和学生端之间通过p2p联机,构成线上VR交互式多媒体教室系统。

关键词：虚拟现实计算机辅助教学课件制作系统 VR 教学系统

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乙二醇浓缩脱水与精制系统操作条件的优化

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计算机与应用化学 2014年第6期31卷 751-754页

作者：赵克品中国石油化工股份有限公司天津分公司研究院天津300271

直接水合成法生产乙二醇过程中,浓缩脱水和精制系统的能耗在整个乙二醇装置中占了很大比重。利用Aspen Plus软件模拟了乙二醇浓缩脱水与精制系统的工业流程。通过工业实际值和模拟值的比较,找到了多个可优化的变量。在此基础上,通过灵... 详细信息

直接水合成法生产乙二醇过程中,浓缩脱水和精制系统的能耗在整个乙二醇装置中占了很大比重。利用Aspen Plus软件模拟了乙二醇浓缩脱水与精制系统的工业流程。通过工业实际值和模拟值的比较,找到了多个可优化的变量。在此基础上,通过灵敏度分析,确定影响该系统能耗的可调变量的调节范围,以系统能耗最低为目标函数,各塔技术指标和产品质量为约束条件,对整个系统进行优化计算,得到了优化的操作条件。在保证产品质量的前提下,采用优化的装置操作条件,系统降低能耗3382 kW,提高系统的操作稳定性。

关键词：乙二醇脱水精制优化流程模拟

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异丙苯氧化产物脱酸工艺研究

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精细石油化工 2021年第6期38卷 14-17页

作者：赵克品中国石油化工股份有限公司天津分公司研究院天津300271

研究了异丙苯氧化产物脱酸工艺,设计了具有一级碱洗+两级逆流水洗工艺的碱洗水洗装置,用于脱除异丙苯氧化产物中的有机酸。考察了碱液量、水量、水洗次数对异丙苯氧化产物中甲酸、乙酸、苯甲酸等有机酸和钠离子的脱除效果,较佳操作条件... 详细信息

研究了异丙苯氧化产物脱酸工艺,设计了具有一级碱洗+两级逆流水洗工艺的碱洗水洗装置,用于脱除异丙苯氧化产物中的有机酸。考察了碱液量、水量、水洗次数对异丙苯氧化产物中甲酸、乙酸、苯甲酸等有机酸和钠离子的脱除效果,较佳操作条件为:氧化液处理量300 L/h,油碱比20∶1、油水比10∶1、一级水洗采用循环水、二级水洗采用新鲜脱盐水,异丙苯氧化液中有机酸含量可由61.93μg/g降至10μg/g以下,钠离子含量由35.9μg/g降至1μg/g以下。

关键词：异丙苯氧化碱洗水洗有机酸钠离子

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纳米氧化镁用于异丙苯催化氧化制过氧化氢异丙苯研究

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精细石油化工 2020年第3期37卷 1-5页

作者：苏跃军赵克品中国石油化工股份有限公司天津分公司天津300270

合成了具有介孔结构的纳米MgO,并对其用于异丙苯选择性氧化制过氧化氢异丙苯(CHP)的催化性能进行了研究。结果表明,纳米MgO在异丙苯选择性氧化制过氧化氢异丙苯过程中表现出良好的催化性能,在MgO添加量为原料质量的1%、空气流速200 mL/... 详细信息

合成了具有介孔结构的纳米MgO,并对其用于异丙苯选择性氧化制过氧化氢异丙苯(CHP)的催化性能进行了研究。结果表明,纳米MgO在异丙苯选择性氧化制过氧化氢异丙苯过程中表现出良好的催化性能,在MgO添加量为原料质量的1%、空气流速200 mL/min、反应温度80℃、常压条件下反应8 h,异丙苯转化率达到30.53%,CHP选择性达到94.78%。

关键词：异丙苯催化氧化过氧化氢异丙苯氧化镁

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异丙苯氧化过程中有机酸的生成及控制

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石油化工 2021年第12期50卷 1238-1245页

作者：张宝忠李军赵克品中国石化天津分公司研究院天津300271

对异丙苯氧化过程中有机酸的生成和控制进行了研究,分析了有机酸生成原理及有机酸对异丙苯氧化反应过程的影响,考察了异丙苯氧化过程中有机酸的主要控制途径。实验结果表明,控制异丙苯氧化反应温度为90~100℃,可减少反应过程中有机酸的... 详细信息

对异丙苯氧化过程中有机酸的生成和控制进行了研究,分析了有机酸生成原理及有机酸对异丙苯氧化反应过程的影响,考察了异丙苯氧化过程中有机酸的主要控制途径。实验结果表明,控制异丙苯氧化反应温度为90~100℃,可减少反应过程中有机酸的生成;反应过程注碱可及时中和生成的有机酸,减少异丙苯氧化反应的副产物;对异丙苯氧化液进行碱洗可有效脱除其中的有机酸和苯酚。采用自制特种高效分离设备,以1%(w)NaOH溶液对异丙苯氧化液进行碱洗,在开启碱液自循环的前提下,在氧化液进料量与碱液注入量的比为15∶1~30∶1范围内,可将异丙苯氧化液中的有机酸含量由53.93×10^(-6)(w)降至20×10^(-6)(w)以下、苯酚含量由349.7×10^(-6)(w)降至60×10^(-6)(w)左右,可满足后续环氧化工段技术要求。

关键词：异丙苯氧化过氧化氢异丙苯有机酸碱洗水洗

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