Develop an efficient and specific AAV-based labeling system for Muller glia in mice

Reprogramming Müller glia (MG) into functional cells is considered a promising therapeutic strategy to treat ocular diseases and vision loss. However, current AAV-based system for MG-tracing was reported to have high leakage in recent studies. Here, we focused on reducing the leakage of AAV-based la...

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Bibliographic Details
Published in:	Scientific reports Vol. 12; no. 1; p. 22410
Main Authors:	Gao, Yanxia, Fang, Kailun, Yan, Zixiang, Zhang, Haiwei, Geng, Guannan, Wu, Weiwei, Xu, Ding, Zhang, Heng, Zhong, Na, Wang, Qifang, Cai, Minqing, Zuo, Erwei, Yang, Hui
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 27-12-2022 Nature Publishing Group Nature Portfolio
Subjects:	631/136/142 631/378/2613/1786 631/378/368 Animal models Animals Cell fate Dependovirus - genetics Genetic Vectors - genetics Glial fibrillary acidic protein Hepatitis Heterogeneous-Nuclear Ribonucleoproteins - genetics Humanities and Social Sciences Humans Labeling Leakage Mice Mueller cells multidisciplinary Neuroglia Polypyrimidine Tract-Binding Protein Post-transcription Regulatory Elements, Transcriptional Science Science (multidisciplinary) Serogroup Serotypes
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Summary:	Reprogramming Müller glia (MG) into functional cells is considered a promising therapeutic strategy to treat ocular diseases and vision loss. However, current AAV-based system for MG-tracing was reported to have high leakage in recent studies. Here, we focused on reducing the leakage of AAV-based labeling systems and found that different AAV serotypes showed a range of efficiency and specificity in labeling MG, leading us to optimize a human GFAP-Cre reporter system packaged in the AAV9 serotype with the woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) removed. The leakage ratio of the AAV9 -hGFAP-Cre- Δ WPRE decreased by an approximate 40-fold compared with the AAV9 -hGFAP-Cre-WPRE labeling system. In addition, we validated the specificity of the AAV-ΔWPRE system for tracing MG reprogramming under Ptbp1-suppression and observed strict non-MG-conversion, similar to previous studies using genetic lineage tracking mouse models. Thus, the AAV9-hGFAP-Cre- Δ WPRE system showed high efficiency and specificity for MG labeling, providing a promising tool for tracing cell fate in vivo.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	2045-2322 2045-2322
DOI:	10.1038/s41598-022-27013-0