Co-editing PINK1 and DJ-1 Genes Via Adeno-Associated Virus-Delivered CRISPR/Cas9 System in Adult Monkey Brain Elicits Classical Parkinsonian Phenotype  

作　　者：Hao Li Shihao Wu Xia Ma Xiao Li Tianlin Cheng Zhifang Chen Jing Wu Longbao Lv Ling Li Liqi Xu Wenchao Wang Yingzhou Hu Haisong Jiang Yong Yin Zilong Qiu Xintian Hu 

机构地区：[1]Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province,Kunming Institute of Zoology,Chinese Academy of Sciences,650223 Kunming,China [2]Institute of Neuroscience,CAS Key Laboratory of Primate Neurobiology,State Key Laboratory of Neuroscience,Chinese Academy of Sciences,Shanghai 200031,China [3]Institutes of Physical Science and Information Technology,Anhui University,Hefei 230601,China [4]Center for Excellence in Brain Science and Intelligence Technology,Chinese Academy of Sciences,Shanghai 200031,China [5]National Resource Center for Non-human Primates,Kunming Primate Research Center,and National Research Facility for Phenotypic and Genetic Analysis of Model Animals(Primate Facility),Kunming Institute of Zoology,Chinese Academy of Sciences,650107 Kunming,China [6]Diagnostic Radiology Department,920 Hospital of the Joint Logistics Support Force of the PLA,Kunming 650032,China [7]Ultrasound diagnosis Department,920 Hospital of the Joint Logistics Support Force of the PLA,Kunming 650032,China [8]Sichuan Provincial People’s Hospital,University of Electronic Science and Technology of China,Chengdu 610072,China [9]Department of Rehabilitation Medicine,The Second People’s Hospital of Yunnan Province,Kunming 650021,China [10]National Clinical Research Center for Aging and Medicine,Huashan Hospital,Fudan University,Shanghai 200433,China

出　　处：《Neuroscience Bulletin》2021年第9期1271-1288,共18页神经科学通报（英文版）

基　　金：This work was supported by the National Key R&D Program of China(2018YFA0801403);the Key-Area Research and Development Program of Guangdong Province(2019B030335001);the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32060200);the National Program for Key Basic Research Projects(973 Program:2015CB755605);the National Natural Science Foundation of China(81471312,81771387,81460352,81500983,31700897,31700910,31800901,31625013,and 91732302);the Applied Basic Research Programs of Science and Technology Commission Foundation of Yunnan Province(2017FB109,2018FB052,2018FB053,2019FA007,and 202001AT070130);Chinese Academy of Sciences"Light of West China"Program,Shanghai Brain-Intelligence Project from Science and Technology Commission of Shanghai Municipality(16JC1420501);Shanghai Municipal Science and Technology Major Project(2018SHZDZX05);Open Large Infrastructure Research of Chinese Academy of Sciences,and China Postdoctoral Science Foundation(2018M631105).

摘　　要：Whether direct manipulation of Parkinson’s disease(PD)risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue.Here,we used an adeno-associated virus serotype 9(AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras(SNs)of two monkey groups:an old group and a middle-aged group.After the operation,the old group exhibited all the classic PD symptoms,including bradykinesia,tremor,and postural instability,accompanied by key pathological hallmarks of PD,such as severe nigral dopaminergic neuron loss(>64%)and evidentα-synuclein pathology in the gene-edited SN.In contrast,the phenotype of their middle-aged counterparts,which also showed clear PD symptoms and pathological hallmarks,were less severe.In addition to the higher final total PD scores and more severe pathological changes,the old group were also more susceptible to gene editing by showing a faster process of PD progression.These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys.Taken together,this system can effectively develop a large number of genetically-edited PD monkeys in a short time(6–10 months),and thus provides a practical transgenic monkey model for future PD studies.

关 键 词：Parkinson’s disease MONKEY Adeno-associated virus-delivered CRISPR/Cas9 PINK1 DJ-1 Parkinsonian phenotype 

分 类 号：R742.5[医药卫生—神经病学与精神病学]