Co-editing PINK1 and DJ-1 Genes Via Adeno-Associated Virus-Delivered CRISPR/Cas9 System in Adult Monkey Brain Elicits Classical Parkinsonian Phenotype
Hao Li1 • Shihao Wu1,2 • Xia Ma1,3 • Xiao Li2 • Tianlin Cheng2 • Zhifang Chen2 • Jing Wu1 • Longbao Lv5 • Ling Li6 • Liqi Xu7 • Wenchao Wang1 • Yingzhou Hu1 • Haisong Jiang8 • Yong Yin9 • Zilong Qiu2,4,10 • Xintian Hu1,4,5
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
Abstract
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 coedit 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 evident a-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 geneticallyedited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.
Keywords
Parkinson’s disease;Monkey;Adeno-associated virus-delivered;CRISPR/Cas9;PINK1;DJ-1;Parkinsonian phenotype
