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| 1 | Effective gene targeting in rabbits using RNA-guided Cas9 nucleases显示文摘 | Dongshan Yang Jie Xu Tianqing Zhu Jianglin Fan Liangxue Lai Jifeng Zhang Y. Eugene Chen | 2014 | Journal of Molecular Cell Biology2014，8，1： | 18 |
| 2 | Generation of RAG 1- and 2-deficient rabbits by embryo microinjection of TALENs显示文摘 | Jun Song Juan Zhong Xiaogang Guo Yongqiang Chen Qingjian zou Jiao Huang Xiaoping Li Quanjun Zhang Zhiwu Jiang Chengcheng Tang Huaqiang Yang Tao Liu Peng Li Duanqing Pei Liangxue Lai | 2013 | Cell Research2013，23，8： | 15 |
| 3 | Species-dependent neuropathology in transgenic SOD' aigs显示文摘 | Huaqiang Yang Guohao Wang Haitao Sun Runzhe Shu Tao Liu Chuan-En Wang Zhaoming Liu Yu Zhao Bentian Zhao Zhen Ouyang Dongshan Yang Jiao Huang Yueling Zhou Shihua Li Xiaodan Jiang Zhicheng Xiao Xiao-Jiang Li Liangxue Lai | 2014 | Cell Research2014，24，4： | 13 |
| 4 | Piglets cloned from induced pluripotent stem cells显示文摘 | Nana Fan Jijun Chen Zhouchun Shang Hongwei Dou Guangzhen Ji Qingjian Zou Lu Wu Lixiazi He Fang Wang Kai Liu Na Liu Jianyong Han Qi Zhou Dengke Pan Dongshan Yang Bentian Zhao Zhen Ouyang Zhaoming Liu Yu Zhao Lin Lin Chongming Zhong Quanlei Wang Shouqi Wang Ying Xu Jing Luan Yu Liang Zhenzhen Yang Jing Li Chunxia Lu Gabor Vajta Ziyi Li Hongsheng Ouyang Huayan Wang Yong Wang Yang Yang Zhonghua Liu Hong Wei Zhidong Luan Miguel A Esteban Hongkui Deng Huanming Yang Duanqing Pei Ning Li Gang Pei Lin Liu Yutao Du Lei Xiao Liangxue Lai | 2013 | Cell Research2013，23，1： | 12 |
| 5 | Genetically Modified Pig Models for Human Diseases显示文摘Genetically modified animal models are important for understanding the pathogenesis of human disease and developing therapeutic strategies.Although genetically modified mice have been widely used to model human diseases,some of these mouse models do not replicate important disease symptoms or pathology.Pigs are more similar to humans than mice in anatomy,physiology,and genome. Thus,pigs are considered to be better animal models to mimic some human diseases.This review describes genetically modified pigs that have been used to model various diseases including neurological,cardiovascular,and diabetic disorders.We also discuss the development in gene modification technology that can facilitate the generation of transgenic pig models for human diseases. | Nana Fan Liangxue Lai | 2013 | Journal of Genetics and Genomics2013，40，2： | 11 |
| 6 | Genome editing in large animals: current status and future prospects显示文摘Large animals(non-human primates,livestock and dogs)are playing important roles in biomedical research,and large livestock animals serve as important sources of meat and milk.The recently developed programmable DNA nucleases have revolutionized the generation of gene-modified large animals that are used for biological and biomedical research.In this review,we briefly introduce the recent advances in nuclease-meditated gene editing tools,and we outline these editing tools’applications in human disease modeling,regenerative medicine and agriculture.Additionally,we provide perspectives regarding the challenges and prospects of the new genome editing technology. | Jianguo Zhao Liangxue Lai Weizhi Ji Qi Zhou | 2019 | National Science Review2019，6，3： | 10 |
| 7 | Genetically humanized pigs exclusively expressing human insulin are generated through custom endonuclease-mediated seamless engineering显示文摘Dear Editor,Type1diabetes(T1D)isa lifelong(chronic)disease and a major health problem throughout the world.This disease can be treatedby either insulin injection or islet transplantation.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. | 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 | 2016 | Journal of Molecular Cell Biology2016，8，2： | 6 |
| 8 | Generation of rat blood vasculature and hematopoietic cells in rat-mouse chimeras by blastocyst complementation显示文摘Interspecies chimera through blastocyst complementation could be an alternative approach to create human organs in animals by using human pluripotent stem cells.A mismatch of the major histocompatibility complex of vascular endothelial cells between the human and host animal will cause graft rejection in the transplanted organs.Therefore,to achieve a transplantable organ in animals without rejection,creation of vascular endothelial cells derived from humans within the organ is necessary.In this study,to explore whether donor xeno-pluripotent stem cells can compensate for blood vasculature in host animals,we generated rat-mouse chimeras by injection of rat embryonic stem cells(rESCs)into mouse blastocysts with deficiency of Flk-1 protein,which is associated with endothelial and hematopoietic cell development.We found that rESCs could differentiate into vascular endothelial and hematopoietic cells in the rat-mouse chimeras.The whole yolk sac(YS)of Flk-1^EGFP/ECFP rat-mouse chimera was full of rat blood vasculature.Rat genes related to vascular endothelial cells,arteries,and veins,blood vessels formation process,as well as hematopoietic cells,were highly expressed in the YS.Our results suggested that rat vascular endothelial cells could undergo proliferation,migration,and self-assembly to form blood vasculature and that hematopoietic cells could differentiate into B cells,T cells,and myeloid cells in rat-mouse chimeras,which was able to rescue early embryonic lethality caused by Flk-1 deficiency in mouse. | Xiaomin Wang Hui Shi Juanjuan Zhou Qingjian Zou Quanjun Zhang Shixue Gou Pengfei Chen Lisha Mou Nana Fan Yangyang Suo Zhen Ouyang Chengdan Lai Quanmei Yan Liangxue Lai | 2020 | Journal of Genetics and Genomics2020，47，5： | 2 |
| 9 | Generation of ApoE deficient dogs via combination of embryo injection of CRISPR/Cas9 with somatic cell nuclear transfer显示文摘Atherosclerotic cardiovascular disease is the leading cause of death in the world which is resulted from complex interactions among multiple genetic and environmental factors(WHO).Atherosclerosis is a chronic inflammatory disease characterized by accumulation of lipids in the arterial wall(Gofman and Lindgren,1950).Tremendous clinical and experimental efforts have | Chong Feng Xiaomin Wang Hui Shi Quanmei Yan Min Zheng Jing Li Quanjun Zhang Yumin Qin Yougang Zhong Jidong Mi Liangxue Lai | 2018 | Journal of Genetics and Genomics2018，45，1： | 2 |
| 10 | Production of transgenic pigs over-expressing theantiviral gene Mx1显示文摘The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interestingcandidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cellnuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs expressapproximately 15–25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was alsomarkedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs withinfluenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profounddecrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-foldreduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found thatthe Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These resultsdemonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate thatthe Mx1 transgene can be harnessed to develop disease-resistant pigs. | Quanmei Yan Huaqiang Yang Dongshan Yang Bentian Zhao Zhen Ouyang Zhaoming Liu Nana Fan Hongsheng Ouyang Weiwang Gu Liangxue Lai | 2014 | Cell Regeneration2014，3，1： | 2 |
| 11 | Efficient and high-fidelity base editor with expanded PAM compatibility for cytidine dinucleotide显示文摘Cytidine base editor(CBE),which is composed of a cytidine deaminase fused to Cas9 nickase,has been widely used to induce C-to-T conversions in a wide range of organisms.However,the targeting scope of current CBEs is largely restricted to protospacer adjacent motif(PAM)sequences containing G,T,or A bases.In this study,we developed a new base editor termed“nNme2-CBE”with excellent PAM compatibility for cytidine dinucleotide,significantly expanding the genome-targeting scope of CBEs.Using nNme2-CBE,targeted editing efficiencies of 29.0%-55.0%and 17.3%—52.5%were generated in human cells and rabbit embryos,respectively.In contrast to conventional nSp-CBE,the nNme2-CBE is a natural high-fidelity base editing platform with minimal DNA off-targeting detected in vivo.Significantly increased efficiency in GC context and precision were determined by combining nNme2Cas9 with rationally engineered cytidine deaminases.In addition,the Founder rabbits with accurate single-base substitutions at Fgf5 gene loci were successfully generated by using the nNme2-CBE system.These novel nNme2-CBEs with expanded PAM compatibility and high fidelity will expand the base editing toolset for efficient gene modification and therapeutic applications. | Zhiquan Liu Siyu Chen Yingqi Jia Huanhuan Shan Mao Chen Yuning Song Liangxue Lai Zhanjun Li | 2021 | Science China(Life Sciences)2021，64，8： | 2 |
| 12 | Vitamin C enhances in vitro and in vivo development of porcine somatic cell nuclear transfer embryos显示文摘 | Yongye Huang Xiaochun Tang Wanhua Xie Yan Zhou Dong Li Yang Zhou Jianguo Zhu Ting Yuan Liangxue Lai Daxin Pang Hongsheng Ouyang | 2011 | Biochemical and Biophysical Research Communications2011，，2： | 1 |
| 13 | Transgenic pig expressing the enhanced green fluorescent protein produced by nuclear transfer using colchicine-treated fibroblasts as donor cells显示文摘 | LAI Liangxue Kwang-wook park Cheong H T | 2002 | Molecular reproduction and development2002，62，： | 1 |
| 14 | Production of cloned transgenic cow expressing omega-3 fatty acids显示文摘 | Xia Wu Hongsheng Ouyang Biao Duan Daxin Pang Li Zhang Ting Yuan Lian Xue Daibang Ni Lei Cheng Shuhua Dong Zhuying Wei Lin Li Ming Yu Qing-Yuan Sun Da-Yuan Chen Liangxue Lai Yifan Dai Guang-Peng Li | 2012 | Transgenic Research2012，，3： | 1 |
| 15 | Mitochondrial distribution and microtubule organization in fertilized and cloned porcine embryos: Implications for developmental potential显示文摘 | Mika Katayama Zhisheng Zhong Liangxue Lai Peter Sutovsky Randall S. Prather Heide Schatten | 2006 | Developmental Biology2006，，1： | 1 |
| 16 | Scriptaid affects histone acetylation and the expression of development-related genes at different stages of porcine somatic cell nuclear transfer embryo during early development显示文摘Although the somatic cell nuclear transfer(SCNT) technique has been used extensively for cloning and generating transgenic pigs,the cloning efficiency is still very low.It has been proposed that the low efficiency of this technique is the result of incomplete epigenetic reprogramming and abnormal gene expression during early embryonic development.In this study,we investigate the effect of Scriptaid,a low-toxicity histone deacetylase inhibitor,on the developmental competence of porcine SCNT embryos.We found that treating SCNT embryos with 500 nmol/L Scriptaid for 15 h after activation significantly enhanced the blastocyst formation rate(27.7%) compared with the untreated group(control)(12.2%,P<0.05).Using an immunofluorescence technique to measure the average fluorescence intensity,we also found that treating SCNT embryos with Scriptaid increased the level of histone acetylation on histone H3 at lysine 14(acH3K14).Furthermore,treating embryos with Scriptaid increased the expression level of three genes that play important roles during embryonic development(Oct4,Klf4 at the blastocyst stage and Nanog at the 4-cell stage).Moreover,the expression level of the apoptosis-related gene Caspase-3 was significantly lower in the Scriptaid-treated SCNT embryos compared with the control SCNT embryos at the 4-cell and blastocyst stages.In conclusion,these results indicate that Scriptaid treatment improves the development and nuclear reprogramming of porcine SCNT embryos. | ZHOU Yan HUANG YongYe XIE WanHua SONG Qi JI Yuan ZHANG YanPing OUYANG HongSheng LAI LiangXue PANG DaXin TANG XiaoChun | 2013 | Chinese Science Bulletin2013，58，18： | 1 |
| 17 | Reprogramming mature terminally differentiated adipocytes to induced pluripotent stem cells显示文摘Mature adipocytes are terminally differentiated somatic cells. Here, we report the successful generation of induced pluripotent stem(i PS) cells from mouse mature adipocytes by forced expression of six transcription factors(Oct4, Sox2, c-Myc, Klf4, Rarc, and Lrh1) with a piggy Bac transposon-based strategy. The resulting i PS cells were pluripotent as evidenced by the fact that they stained positive for alkaline phosphatase, expressed high levels of key pluripotency markers including Oct4, Nanog, and SSEA1,and remained pluripotent on a 2i media. In vitro differentiation of the i PS cells showed that the cell derivatives of all three germ layers could be readily obtained through formation of embryoid bodies. Most importantly, these adipocytederived i PS cells were capable of producing chimera with high frequencies when reintroduced into early-stage embryos and transmitted through the germ line. This study demonstrates that the new six-factor reprogramming technology facilitates the reset of the terminally differentiated adipocytes to the ground state of pluripotency, enabling us to fully explore the potential of mature adipocytes as a viable cell source for regenerative medicine. | Tao Nie Wei Deng Xuefei Gao Wei Sun Xiaoyan Hui Hong Song Dajiang Qin Aimin Xu Peng Li Pentao Liu Liangxue Lai Donghai Wu | 2015 | Science Bulletin2015，60，20： | 1 |
| 18 | In vitro development of preimplantation porcine nuclear transfer embryos cultured in different media and gas atmospheres显示文摘 | Gi-Sun Im Liangxue Lai Zhonghua Liu Yanhong Hao David Wax Aaron Bonk Randall S Prather | 2003 | Theriogenology2003，，6： | 1 |
| 19 | Nuclear reprogramming by nuclear transplantation and defined transcription factors显示文摘In the past ten years,great breakthroughs have been achieved in the nuclear reprogramming area. It has been demonstrated that highly differentiated somatic cell genome could be reprogrammed to a pluripotent state,which indicates that differentiated cell fate is not irreversible. Nuclear transplantation and induced pluripotent stem (iPS) cell generation are the two major approaches to inducing reprogramming of differentiated somatic cell genome. In the present review,we will summarize the recent progress of nuclear reprogramming and further discuss the potential to generate patient specific pluripotent stem cells from differentiated somatic cells for therapeutic purpose. | WANG YiXuan LIU Sheng LAI LiangXue GAO ShaoRong | 2009 | Chinese Science Bulletin2009，54，1： | 1 |
| 20 | Generation of gene-target dogs using CRISPR/Cas9 system显示文摘Dear Editor,Dogs(Canis familiaris)serve as human companions and are raised to herd livestock,aid hunters,guard homes,perform police and rescue work,and guide the blind.Dogs exhibit close similarities to humans in terms of metabolic,physiological,and anatomical characteristics,and thus are ideal genetic and clinical models to study human diseases(Tsai et al.,2007).Gene target technology is a powerful tool to create new strains of animals with favorable traits.However,thus far,gene-target dogs have not been developed due to their unique species-specific reproductive characteristics,which limits the applications of dogs especially in the field of biomedical research.Recently,clustered regularly interspaced short palindromic repeats(CRISPRs)/CRISPR-associated(Cas)9 system was applied to edit specific genes with a high efficiency(Cong et al.,2013;Mali et al.,2013).Here we attempt to explore the feasibility of producing gene knockout(KO)dogs by using this technology.Beagle dog,the most widely used breed in biomedical research,was used as our animal model.Myostatin(MSTN)was chosen as the first gene of interest. | Qingjian Zou Xiaomin Wang Yunzhong Liu Zhen Ouyang Haibin Long Shu Wei Jige Xin Bentian Zhao Sisi Lai Jun Shen Qingchun Ni Huaqiang Yang Huilin Zhong Li Li Minhua Hu Quanjun Zhang Zhidong Zhou Jiaxin He Quanmei Yan Nana Fan Yu Zhao Zhaoming Liu Lin Guo Jiao Huang Guanguan Zhang Jun Ying Liangxue Lai Xiang Gao | 2015 | Journal of Molecular Cell Biology2015，7，6： | 1 |
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