@article{92c7913136784a28826188588ae0c80d,
title = "Critical examination of Ptbp1-mediated glia-to-neuron conversion in the mouse retina",
abstract = "Reprogramming glial cells to convert them into neurons represents a potential therapeutic strategy that could repair damaged neural circuits and restore function. Recent studies show that downregulation of the RNA-binding protein PTBP1 leads to one-step conversion of M{\"u}ller glia (MG) into retinal ganglion cells (RGCs) with a high efficiency. However, the original study did not perform fate-mapping experiments to confirm MG-to-RGC conversion after Ptbp1 downregulation. To address the fundamental question of whether Ptbp1 downregulation can convert MG into RGCs in the mouse retina, we perform fate-mapping experiments to lineage trace MG independent of the adeno-associated virus (AAV)-mediated labeling system. Here, we report that Ptbp1 downregulation by CRISPR-CasRx or small hairpin RNA is insufficient to convert MG to RGCs. The original conclusion of MG-to-RGC conversion is due to leaky labeling of endogenous RGCs. Our results emphasize the importance of using stringent fate mapping to determine glia-to-neuron conversion in cell reprogramming research.",
keywords = "CP: Neuroscience, CP: Stem cell research, M{\"u}ller glia, fate mapping, glia-to-neuron conversion, lineage tracing, reprogramming, retinal ganglion cells",
author = "Ye Xie and Jing Zhou and Bo Chen",
note = "Funding Information: We thank Dr. Xiang-Dong Fu (University of California, San Diego), for providing AAV-CMV-LSL-RFP and AAV-CMV-LSL-RFP-shPtbp1 plasmids and technical advice, and the flow cytometry CoRE (Icahn School of Medicine at Mount Sinai) for technical help with MG purification. This work was supported by National Institutes of Health grants R01 EY024986 and R01 EY028921 , an unrestricted challenge grant from Research to Prevent Blindness , the New York Eye and Ear Infirmary Foundation , and The Harold W. McGraw Jr. Family Foundation for Vision Research . Funding Information: We thank Dr. Xiang-Dong Fu (University of California, San Diego), for providing AAV-CMV-LSL-RFP and AAV-CMV-LSL-RFP-shPtbp1 plasmids and technical advice, and the flow cytometry CoRE (Icahn School of Medicine at Mount Sinai) for technical help with MG purification. This work was supported by National Institutes of Health grants R01 EY024986 and R01 EY028921, an unrestricted challenge grant from Research to Prevent Blindness, the New York Eye and Ear Infirmary Foundation, and The Harold W. McGraw Jr. Family Foundation for Vision Research. Y.X. and J.Z. designed and performed experiments and analyzed data. B.C. conceived the project, designed experiments, analyzed data, and supervised the project. Y.X. and B.C. wrote the paper. All authors have read and approved the paper. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = jun,
day = "14",
doi = "10.1016/j.celrep.2022.110960",
language = "English",
volume = "39",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "11",
}