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Cell reprogramming therapy for Parkinson’s disease

Neural Regeneration Research 2024年第11期19卷 2444-2455页

作者： Wenjing Dong Shuyi liu shangang li Zhengbo Wang State Key Laboratory of Primate Biomedical Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunmingYunnan ProvinceChina Yunnan Key Laboratory of Primate Biomedical Research KunmingYunnan ProvinceChina

Parkinson’s disease is typically characterized by the progressive loss of dopaminergic neurons in the substantia nigra pars *** studies have been performed based on the supplementation of lost dopaminergic neurons to treat Parkinson’s *** initial strategy for cell replacement therapy used human fetal ventral midbrain and human embryonic stem cells to treat Parkinson’s disease,which could substantially alleviate the symptoms of Parkinson’s disease in clinical ***,ethical issues and tumor formation were limitations of its clinical *** pluripotent stem cells can be acquired without sacrificing human embryos,which eliminates the huge ethical barriers of human stem cell *** widely considered neuronal regeneration strategy is to directly reprogram fibroblasts and astrocytes into neurons,without the need for intermediate proliferation states,thus avoiding issues of immune rejection and tumor *** induced pluripotent stem cells and direct reprogramming of lineage cells have shown promising results in the treatment of Parkinson’s ***,there are also ethical concerns and the risk of tumor formation that need to be *** review highlights the current application status of cell reprogramming in the treatment of Parkinson’s disease,focusing on the use of induced pluripotent stem cells in cell replacement therapy,including preclinical animal models and progress in clinical *** review also discusses the advancements in direct reprogramming of lineage cells in the treatment of Parkinson’s disease,as well as the controversy surrounding in vivo *** findings suggest that cell reprogramming may hold great promise as a potential strategy for treating Parkinson’s disease.

关键词： animal models astrocytes autologous cell reprogramming cell therapy direct lineage reprogramming dopaminergic neurons induced pluripotent stem cells non-human primates Parkinson’s disease

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Identification of microsatellites in cattle unigenes

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Journal of Genetics and Genomics 2008年第5期35卷 261-266页

作者： Qiuliang Yan Yinghan Zhang Hongbin li Caihong Wei lili Niu Shan Guan shangang li lixin Du College of Animal Science and Technology NorthwestA ＆ F University Yangling 712100 China National Center for Molecular Genetics and Breeding of Animal Institute of Animal Sciences Chinese Academy of Agricultural Sciences Beijing 100094 China Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 China

To identify EST-SSR molecular markers, 41,986 cattle UniGene sequences from NCBI were mined for analyzing SSRs. A total of 1,831 SSRs were identified from 1,666 ESTs, which represented an average density of 19.88 kb per SSR. The frequency of EST-SSRs was 4.0%. The dinucleotide repeat motif was the most abundant SSR, accounting for 54%, followed by 22%, 13%, 7% and 4%, respectively, for tri-, hexa-, penta- and tetra-nucleotide repeats. Depending upon the length of the repeat unit, the length of microsatellites varied from 14 to 86 bp. Among the di- and tri-nucleotide repeats, AC/TG （57%） and AGC （12%） were the most abundant type. Annotation of EST-SSRs was also carried out. Three hundred primer pairs were randomly designed using Prime Premier 5.0 program and Oligo 5.0 for further experimental validation.

关键词： cattle microsatellite expressed sequence tags EST-SSR

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纳米粒子在CRISPR/Cas9基因治疗中的应用

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生物工程学报 2022年第6期38卷 2087-2104页

作者：马跃邓莉李善刚昆明理工大学灵长类转化医学院省部共建非人灵长类生物医学国家重点实验室云南中科灵长类生物医学实验室云南昆明650000

CRISPR/Cas9基因编辑技术已成为基因治疗领域最有前景的工具。在临床应用中,对CRISPR/Cas9进行安全有效的递送一直是亟待解决的问题。纳米粒子,如脂基纳米粒子、聚合物纳米粒子、纳米金颗粒以及生物膜类纳米粒子等,因其生物相容性、安... 详细信息

CRISPR/Cas9基因编辑技术已成为基因治疗领域最有前景的工具。在临床应用中,对CRISPR/Cas9进行安全有效的递送一直是亟待解决的问题。纳米粒子,如脂基纳米粒子、聚合物纳米粒子、纳米金颗粒以及生物膜类纳米粒子等,因其生物相容性、安全性和可设计性等特点有望为基因治疗带来新的突破。文中首先对纳米粒子的特性和基因治疗中CRISPR/Cas9的发展进行了概述,然后详细归纳了纳米粒子在递送不同形式的CRISPR/Cas9中的应用,最后对纳米粒子介导的基因治疗的递送在未来面临的挑战和安全性等方面作出总结论述。

关键词： CRISPR/Cas9 基因治疗非病毒载体纳米粒子

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Interspecies embryo transfer between rhesus and cynomolgus monkeys

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Journal of Genetics and Genomics 2020年第6期47卷 333-336页

作者： Ting Zhang Yu Kang li li Yin Zhou Xinglong Chen Yan Zhuo Zifan li Hong Wang Yuyu Niu Weizhi Ji shangang li Yongchang Chen Yunnan Key Laboratory of Primate Biomedicine Research Institute of Primate Translational MedicineKunming University of Science and TechnologyKunming650500China Faculty of Environmental Science and Engineering Kunming University of Science and TechnologyKunming650500China Department of Pediatrics The First People's Hospital of Yunnan Province and The Affiliated Hospital of Kunming University of Science and TechnologyKunming650032China

Nonhuman primates(NHPs)present significant advantages over rodents for physiological and pharmacological investigations because of their higher genetic and physiological similarity to humans.

关键词： primates transfer similarity

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DMD 的肌膜损伤机制及其修复治疗的研究进展

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中国比较医学杂志 2022年第6期32卷 93-99页

作者：赵治鹏李善刚昆明理工大学灵长类转化医学研究院昆明650500 省部共建非人灵长类生物医学国家重点实验室昆明650500

杜氏肌营养不良症(Duchenne muscular dystrophy,DMD)是一种严重的致死性肌肉疾病,定位于肌膜的抗肌萎缩蛋白dystrophin功能缺失是其致病因素。目前DMD研究的热点聚焦于如何对疾病进行治疗,其中膜功能的恢复已经成为临床治疗的密切关注... 详细信息

杜氏肌营养不良症(Duchenne muscular dystrophy,DMD)是一种严重的致死性肌肉疾病,定位于肌膜的抗肌萎缩蛋白dystrophin功能缺失是其致病因素。目前DMD研究的热点聚焦于如何对疾病进行治疗,其中膜功能的恢复已经成为临床治疗的密切关注点。本综述将着重总结dystrophin异常引起肌膜损伤的机制及膜修复治疗的研究进展,通过从肌膜的角度理解DMD的病理机制来促进干预治疗的研究进程。

关键词：杜氏肌营养不良症肌膜损伤肌膜修复治疗

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CRISPR/Cas9-mediated mutation of asparagine-linked glycosylation 13 transcript variant 1 causes epilepsy in mice

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Journal of Bio-X Research 2020年第1期3卷 6-12页

作者： Yu Baoli Zhu Geng li shangang Chen Xuejin Fang Qian Zhang Yan liu Yixun li Shengtian Wu Ji Renji Hospital Shanghai Jiaotong University School of Medicine Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders(Ministry of Education)Bio-X InstitutesShanghai Jiao Tong UniversityShanghai Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education Ningxia Medical UniversityYinchuanNingxia Hui Autonomous Region Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders(Ministry of Education) Bio-X InstitutesShanghai Jiao Tong University Department of Biomedical Engineering School of Medical InstrumentShanghai University of Medicine&Health Sciences Department of Laboratory Animal Sciences Shanghai Jiao Tong University School of MedicineShanghai Institute of Biophysics Chinese Academy of Sciences State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijingChina

Objective:The asparagine-linked glycosylation 13 homolog(Alg13)has been identified as causative for congenital disorders of glycosylation type I with *** aim of this study was to determine whether mice carrying a mutated version of Alg13 could be used as a model for epileptic encephalopathies or congenital disorders of glycosylation type ***:A model of epileptic encephalopathy was established in C57BL/6 mice by introducing mutations in Alg13 via the clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(CRISPR/Cas9)*** surgical procedures were approved by the Institutional Animal Care and Use Committee of Shanghai Jiao Tong University(A2016084)on October 8,***:Mice with 3 different mutations,Alg13^(-54nt),Alg13^(-5nt)and Alg13^(-4nt),all of which are located in Alg13 transcript variant 1,were *** Alg13^(-5nt)mice exhibited spontaneous seizures similar to patients with Alg13 mutations,suggesting that they could be used as a model for *** blot analysis demonstrated that Alg13^(-5nt)mice had lower levels of Alg13 expression than wild-type *** observations showed that two of the 17 Alg13^(-5nt)mice had stage 5 seizures involving jumping and falling,while 12 had stage 3 seizures with head ***:The Alg13 mouse model provides an outstanding tool for studying epileptic encephalopathies and investigating different aspects of defects in glycosylation or other post-translational modification that cannot be assessed in patients or cell culture systems.

关键词： Alg13 CRISPR/Cas9 epilepsy mouse mutant

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基于CRISPR/Cas9系统和增强ssODN重组建立tau-V337M小鼠模型

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生物工程学报 2023年第7期39卷 3003-3014页

作者：陈丽娇邓莉孙文杰刘杰张婷李善刚昆明理工大学灵长类转化医学研究院省部共建非人灵长类生物医学国家重点实验室云南昆明650500 云南中科灵长类生物医学重点实验室云南昆明650500

利用CRISPR/Cas9基因编辑技术建立tau-V337M突变的阿尔茨海默病(Alzheimer’s disease,AD)小鼠模型。通过设计和体外合成单向导RNAs(single guide RNAs,sgRNA)及单链寡核苷酸(single-stranded oligonucleotides,ssODN),将sgRNA、Cas9蛋白、ssODN注射到小鼠受精卵内,利用DNA切割和重组产生突变。为了提高重组效率,又在注射时添加Rad51蛋白。使用自然交配的雌鼠作为受体,将2细胞期的编辑胚胎进行单侧输卵管移植。研究发现通过添加Rad51蛋白可以获得较高的突变效率,在F0小鼠中获得了tau-V337M小鼠并进行扩繁,F0代tau-V337M小鼠可以将突变遗传给F1代。综上所述,本研究利用Cas9、ssODN和Rad51成功建立了首个tau-V337M基因位点突变的小鼠模型,为AD的研究和点突变模型制作提供了模型和方法基础。

关键词：阿尔茨海默症 tau-V337M Cas9 DNA重组单侧输卵管移植 Rad51

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Extensive humoral immune response to AAVs and Cas proteins in nonhuman primates

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Science Bulletin 2021年第20期66卷 2061-2064,M0003页

作者： Puhao Xiao Raoxian Bai Ting Zhang Ruo Wu lijiao Chen Yu Hou Bin Shen Yuyu Niu shangang li Weizhi Ji Yongchang Chen State Key Laboratory of Primate Biomedical Research/Institute of Primate Translational Medicine Kunming University of Science and TechnologyKunming 650500China State Key Laboratory of Reproductive Medicine Department of Histology and EmbryologyNanjing Medical UniversityNanjing 210029China

Gene therapy is expected to fundamentally correct mutant genes to cure genetic diseases in *** a delivery system for gene therapy,adenovirus-associated virus(AAV)vectors have achieved positive results in clinical and ... 详细信息

关键词： AAV 体液免疫反应体液免疫应答非人灵长类基因治疗基因编辑恒河猴

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CRISPR/Cas9系统诱导DNA断裂的修复机制研究进展

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中国细胞生物学学报 2022年第3期44卷 500-511页

作者：杨丽芸陈丽娇李善刚昆明理工大学灵长类转化医学研究院省部共建非人灵长类生物医学国家重点实验室云南中科灵长类生物医学重点实验室昆明650500

近年来,聚类规则间隔的短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)及其相关蛋白(CRISPR-associated protein 9,Cas9)介导的基因编辑技术已广泛应用在生物学、基础医学等研究领域中,通过该技术构建的动物疾病模型推动了对发病机制的探索和基因治疗等方面的研究进展。Cas9核酸酶切割与随后的同源重组(HDR)或非同源末端连接(NHEJ)共同完成基因编辑工作,但是HDR发生的效率较低限制了其应用。因此对NHEJ和HDR修复方式的机制研究对于提高基因编辑的精准性及高效性十分重要。该文主要对CRISPR/Cas9系统应用过程中NHEJ和HDR修复系统的发生机制进行总结并结合相关研究进展进行了展望,为高效利用CRISPR/Cas9技术的研究提供了思路。

关键词： CRISPR/Cas9 DSB 同源重组非同源末端连接疾病模型基因治疗

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Transgene expression of enhanced green fluorescent protein in cloned rabbits generated from in vitro-transfected adult fibroblasts

Transgene expression of enhanced green fluorescent protein i...

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第九届中国实验动物科学年会（2010新疆）

作者： shangang li Yi Guo Jianjun Shi Chunguang Yin Fengying Xing lingyang Xu Chuanshan Zhang Tao liu Yao li Hongbin li lixin Du Xuejin Chen

Objective live rabbits have previously been generatedthrough nuclear transfer using adult cells as nuclear *** demonstrated in this study that transfected adult rabbit fibroblasts are also capableof supporting full-term *** fibroblasts were transfected with a pEGFP-C1 plasmid using lipofectamine TM 2000,and the transgenic cells were derived from conditioned *** transgenic fibroblasts were cultured until confluent and then serum-starved prior to be used as nuclear *** nuclear transfer and activation,22%(12/55) of the transgenic cloned embryos developed to the blastocyst stage.A total of 114 embryos at the 4- to 8-cell stage were transferred to the oviducts of 8 pseudo-pregnant mothers;5 of these animals became pregnant,and 3 of the 5 mother rabbits carried the pregnancy to term. Caesarean section was performed on the 3 pregnant mothers,yielding 4 kits,one of which has survived for more than 9 *** fluorescence could be detected in the toenails of the living cloned rabbit and the offspring from the living cloned rabbit under ultraviolet *** analyses confirmed that all 4 cloned rabbits were genetically identical to the transgenic donor cells,and that they all carried the EGFP *** present study demonstrated that transgenic rabbits can be generated through nuclear *** results may facilitate future developments in the genetic engineering of rabbits.

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