Striatonigral distribution of a fluorescent reporter following intracerebral delivery of genome editors

Frontiers in Bioengineering and Biotechnology

Bioengineering and Biotechnology

Krishanu Saha¹  Yuyuan Wang²  Shaoqin Gong²  Viktoriya Bondarenko³  Jeanette M. Metzger³  Jesi Felton³  Samuel S. Neuman³  Marina E. Emborg⁴  Jon E. Levine⁵ 

[1] Department of Biomedical Engineering, University of Wisconsin–Madison, Madison, WI, United States;Wisconsin Institute for Discovery, University of Wisconsin–Madison, Madison, WI, United States;Department of Ophthalmology and Visual Sciences, University of Wisconsin–Madison, Madison, WI, United States;Department of Biomedical Engineering, University of Wisconsin–Madison, Madison, WI, United States;Wisconsin Institute for Discovery, University of Wisconsin–Madison, Madison, WI, United States;Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, United States;Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, United States;Department of Medical Physics, University of Wisconsin-Madison, Madison, WI, United States;Wisconsin National Primate Research Center, University of Wisconsin-Madison, Madison, WI, United States;Department of Neuroscience, University of Wisconsin–Madison, Madison, WI, United States;

关键词: nanomedicine;    CRISPR;    viral vectors;    Parkinson’s disease;    neural networks;    substantia nigra;    gene editing;   

DOI  :  10.3389/fbioe.2023.1237613

 received in 2023-06-09, accepted in 2023-07-18,  发布年份 2023

来源: Frontiers

【 摘 要 】

Introduction: Targeted gene editing is proposed as a therapeutic approach for numerous disorders, including neurological diseases. As the brain is organized into neural networks, it is critical to understand how anatomically connected structures are affected by genome editing. For example, neurons in the substantia nigra pars compacta (SNpc) project to the striatum, and the striatum contains neurons that project to the substantia nigra pars reticulata (SNpr).Methods: Here, we report the effect of injecting genome editors into the striatum of Ai14 reporter mice, which have a LoxP-flanked stop cassette that prevents expression of the red fluorescent protein tdTomato. Two weeks following intracerebral delivery of either synthetic nanocapsules (NCs) containing CRISPR ribonucleoprotein targeting the tdTomato stop cassette or adeno-associated virus (AAV) vectors expressing Cre recombinase, the brains were collected, and the presence of tdTomato was assessed in both the striatum and SN.Results: TdTomato expression was observed at the injection site in both the NC- and AAV-treated groups and typically colocalized with the neuronal marker NeuN. In the SN, tdTomato-positive fibers were present in the pars reticulata, and SNpr area expressing tdTomato correlated with the size of the striatal genome edited area.Conclusion: These results demonstrate in vivo anterograde axonal transport of reporter gene protein products to the SNpr following neuronal genome editing in the striatum.

【 授权许可】

Unknown   
Copyright © 2023 Neuman, Metzger, Bondarenko, Wang, Felton, Levine, Saha, Gong and Emborg.