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M\u00fcller glia (MG), the primary glial cell type in the retina, exhibit extraordinary regenerative abilities in zebrafish, proliferating and differentiating into neurons post-injury. However, the regenerative potential of mouse MG is limited by their inherent inability to re-enter the cell cycle, constrained by high levels of the cell cycle inhibitor p27\n                    <jats:sup>Kip1<\/jats:sup>\n                    and low levels of cyclin D1. Here, we report a method to drive robust MG proliferation by adeno-associated virus (AAV)-mediated cyclin D1 overexpression and p27\n                    <jats:sup>Kip1<\/jats:sup>\n                    knockdown. MG proliferation induced by this dual targeting vector was self-limiting, as MG re-entered cell cycle only once. As shown by single-cell RNA-sequencing, cell cycle reactivation led to suppression of interferon signaling, activation of reactive gliosis, and downregulation of glial genes in MG. Over time, the majority of the MG daughter cells retained the glial fate, resulting in an expanded MG pool. Interestingly, about 1% MG daughter cells expressed markers for retinal interneurons, suggesting latent neurogenic potential in a small MG subset. By establishing a safe, controlled method to promote MG proliferation in vivo while preserving retinal integrity, this work provides a valuable tool for combinatorial therapies integrating neurogenic stimuli to promote neuron regeneration.\n                  <\/jats:p>","DOI":"10.7554\/elife.100904","type":"journal-article","created":{"date-parts":[[2024,8,28]],"date-time":"2024-08-28T14:25:30Z","timestamp":1724855130000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.100904","source":"Crossref","is-referenced-by-count":8,"title":["Simultaneous cyclin D1 overexpression and p27kip1 knockdown enable robust M\u00fcller glia cell cycle reactivation in uninjured mouse retina"],"prefix":"10.7554","volume":"13","author":[{"given":"Zhifei","family":"Wu","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03q8dnn23","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Biomedical Sciences and Tung Biomedical Sciences Centre, City University of Hong 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