{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T11:39:54Z","timestamp":1792582794993,"version":"4.5.0"},"reference-count":28,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T00:00:00Z","timestamp":1751328000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T00:00:00Z","timestamp":1751328000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>Optogenetic gene therapy is a promising mutation-independent treatment that aims to restore visual perception in patients blinded by retinal diseases that cause photoreceptor degeneration. Still, low sensitivity or slow kinetics of currently utilized optogenetic proteins limit the efficacy of such approaches. Here, we evaluated the therapeutic potential of three channelrhodopsin variants: ChRmine, from the algae <jats:italic>Rhodomonas lens<\/jats:italic>, ChRmine-T119A, a faster-closing ChRmine variant, and ChroME2s, a second-generation Chronos-based opsin.We expressed these opsins in retinal ganglion cells of <jats:italic>rd1<\/jats:italic> mice, a model of severe retinal degeneration. Single cell electrophysiology demonstrates opsin\u2019s large sensitivity to a range of light intensities as well as opsin-expressing retinal ganglion cells generated action potentials in response to light stimulation. Behavioral tests showed ChRmine-T119A\u2019s efficacy at 360\u00a0lux compared to unmodified ChRmine and ChroME2s. ChRmine and ChroME2s did restore light perception at higher light intensities. Additionally, our dose\u2013response study with ChRmine-T119A revealed that lower viral titers were more effective at restoring light sensitivity. Our study demonstrates that these ChRmine- and ChroME-based opsins can enhance vision in late-stage blinding diseases.<\/jats:p>","DOI":"10.1038\/s41598-025-04286-9","type":"journal-article","created":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T16:39:41Z","timestamp":1751387981000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Optogenetic restoration of high-sensitivity vision using ChRmine- and ChroME-based channelrhodopsins"],"prefix":"10.1038","volume":"15","author":[{"given":"Victoria C.","family":"Fong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beatrice M.","family":"Le","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonia","family":"Stefanov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivian","family":"Lee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seohyun","family":"Park","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adithya","family":"Sivakumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabrina","family":"Spatny","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meike","family":"Visel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. Rowland","family":"Taylor","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen G.","family":"Brohawn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John G.","family":"Flannery","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,1]]},"reference":[{"key":"4286_CR1","doi-asserted-by":"publisher","first-page":"846","DOI":"10.1002\/ajmg.c.31842","volume":"184","author":"CA Ku","year":"2020","unstructured":"Ku, C. A. & Pennesi, M. E. The new landscape of retinal gene therapy. Am. J. Med. Genet. C Semin. Med. Genet. 184, 846\u2013859 (2020).","journal-title":"Am. J. Med. Genet. C Semin. Med. Genet."},{"key":"4286_CR2","doi-asserted-by":"publisher","DOI":"10.1016\/j.preteyeres.2021.100975","volume":"86","author":"C Botto","year":"2022","unstructured":"Botto, C. et al. Early and late stage gene therapy interventions for inherited retinal degenerations. Prog. Retin. Eye Res. 86, 100975 (2022).","journal-title":"Prog. Retin. Eye Res."},{"key":"4286_CR3","doi-asserted-by":"publisher","DOI":"10.1101\/cshperspect.a041304","volume":"13","author":"A Stefanov","year":"2023","unstructured":"Stefanov, A. & Flannery, J. G. A systematic review of optogenetic vision restoration: History, challenges, and new inventions from bench to bedside. Cold Spring. Harb. Perspect. Med. 13, a041154 (2023).","journal-title":"Cold Spring. Harb. Perspect. Med."},{"key":"4286_CR4","doi-asserted-by":"publisher","first-page":"1099348","DOI":"10.3389\/fnana.2022.1099348","volume":"16","author":"RL Pfeiffer","year":"2022","unstructured":"Pfeiffer, R. L. & Jones, B. W. Current perspective on retinal remodeling: Implications for therapeutics. Front. Neuroanat. 16, 1099348 (2022).","journal-title":"Front. Neuroanat."},{"key":"4286_CR5","doi-asserted-by":"publisher","first-page":"525","DOI":"10.1007\/978-1-4419-1399-9_60","volume":"664","author":"T Puthussery","year":"2010","unstructured":"Puthussery, T. & Taylor, W. R. Functional changes in inner retinal neurons in animal models of photoreceptor degeneration. Adv. Exp. Med. Biol. 664, 525\u2013532 (2010).","journal-title":"Adv. Exp. Med. Biol."},{"key":"4286_CR6","doi-asserted-by":"publisher","first-page":"14282","DOI":"10.1523\/JNEUROSCI.4968-08.2008","volume":"28","author":"F Mazzoni","year":"2008","unstructured":"Mazzoni, F., Novelli, E. & Strettoi, E. Retinal ganglion cells survive and maintain normal dendritic morphology in a mouse model of inherited photoreceptor degeneration. J. Neurosci. 28, 14282\u201314292 (2008).","journal-title":"J. Neurosci."},{"key":"4286_CR7","doi-asserted-by":"publisher","first-page":"1429","DOI":"10.1016\/j.stemcr.2018.04.010","volume":"10","author":"RA Barker","year":"2018","unstructured":"Barker, R. A. et al. The challenges of first-in-human stem cell clinical trials: What does this mean for ethics and institutional review boards?. Stem. Cell Rep. 10, 1429\u20131431 (2018).","journal-title":"Stem. Cell Rep."},{"key":"4286_CR8","doi-asserted-by":"publisher","first-page":"1223","DOI":"10.1038\/s41591-021-01351-4","volume":"27","author":"JA Sahel","year":"2021","unstructured":"Sahel, J. A. et al. Partial recovery of visual function in a blind patient after optogenetic therapy. Nat. Med. 27, 1223\u20131229 (2021).","journal-title":"Nat. Med."},{"key":"4286_CR9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0098924","volume":"9","author":"ZH Pan","year":"2014","unstructured":"Pan, Z. H., Ganjawala, T. H., Lu, Q., Ivanova, E. & Zhang, Z. ChR2 mutants at L132 and T159 with improved operational light sensitivity for vision restoration. PLoS ONE 9, e98924 (2014).","journal-title":"PLoS ONE"},{"key":"4286_CR10","doi-asserted-by":"publisher","first-page":"1139","DOI":"10.1016\/j.neuron.2022.01.008","volume":"110","author":"S Sridharan","year":"2022","unstructured":"Sridharan, S. et al. High-performance microbial opsins for spatially and temporally precise perturbations of large neuronal networks. Neuron 110, 1139-1155.e6 (2022).","journal-title":"Neuron"},{"key":"4286_CR11","doi-asserted-by":"publisher","first-page":"11642","DOI":"10.1038\/s41598-024-62558-2","volume":"14","author":"H Bansal","year":"2024","unstructured":"Bansal, H., Pyari, G. & Roy, S. Theoretical prediction of broadband ambient light optogenetic vision restoration with ChRmine and its mutants. Sci. Rep. 14, 11642 (2024).","journal-title":"Sci. Rep."},{"key":"4286_CR12","doi-asserted-by":"publisher","first-page":"4842","DOI":"10.1038\/s41467-022-32441-7","volume":"13","author":"K Tucker","year":"2022","unstructured":"Tucker, K., Sridharan, S., Adesnik, H. & Brohawn, S. G. Cryo-EM structures of the channelrhodopsin ChRmine in lipid nanodiscs. Nat. Commun. 13, 4842 (2022).","journal-title":"Nat. Commun."},{"key":"4286_CR13","doi-asserted-by":"publisher","first-page":"394","DOI":"10.1001\/archopht.1974.03900060406015","volume":"91","author":"MM LaVail","year":"1974","unstructured":"LaVail, M. M. & Sidman, R. L. C57BL\/6J Mice with inherited retinal degeneration. Arch. Ophthalmol. 91, 394\u2013400 (1974).","journal-title":"Arch. Ophthalmol."},{"key":"4286_CR14","doi-asserted-by":"publisher","first-page":"2019","DOI":"10.1038\/s41598-024-52391-y","volume":"14","author":"L Zhang","year":"2024","unstructured":"Zhang, L. et al. The temporal progression of retinal degeneration and early-stage idebenone treatment in the Pde6b rd1\/rd1 mouse model of retinal dystrophy. Sci. Rep. 14, 2019 (2024).","journal-title":"Sci. Rep."},{"issue":"385","key":"4286_CR15","first-page":"397","volume":"25","author":"ST Lim","year":"2000","unstructured":"Lim, S. T., Antonucci, D. E., Scannevin, R. H. & Trimmer, J. S. A. Novel targeting signal for proximal clustering of the Kv2.1 K+ channel in hippocampal neurons. Neuron 25(385), 397 (2000).","journal-title":"Neuron"},{"key":"4286_CR16","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41434-022-00380-z","volume":"30","author":"B Nieuwenhuis","year":"2023","unstructured":"Nieuwenhuis, B. et al. Improving adeno-associated viral (AAV) vector-mediated transgene expression in retinal ganglion cells: Comparison of five promoters. Gene. Ther. 30, 1\u201317 (2023).","journal-title":"Gene. Ther."},{"key":"4286_CR17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0018556","volume":"6","author":"JH Kim","year":"2011","unstructured":"Kim, J. H. et al. High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish mice. PLoS ONE 6, e18556 (2011).","journal-title":"PLoS ONE"},{"key":"4286_CR18","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1038\/s41467-023-36467-3","volume":"14","author":"A Luo","year":"2023","unstructured":"Luo, A. et al. H2B ubiquitination recruits FACT to maintain a stable altered nucleosome state for transcriptional activation. Nat. Commun. 14, 741 (2023).","journal-title":"Nat. Commun."},{"key":"4286_CR19","doi-asserted-by":"publisher","first-page":"672","DOI":"10.1016\/j.cell.2022.01.007","volume":"185","author":"KE Kishi","year":"2022","unstructured":"Kishi, K. E. et al. Structural basis for channel conduction in the pump-like channelrhodopsin ChRmine. Cell 185, 672\u2013689 (2022).","journal-title":"Cell"},{"key":"4286_CR20","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1167\/7.14.7","volume":"7","author":"G Wang","year":"2007","unstructured":"Wang, G. et al. The sensitivity of light-evoked responses of retinal ganglion cells is decreased in nitric oxide synthase gene knockout mice. J. Vis. 7, 7 (2007).","journal-title":"J. Vis."},{"key":"4286_CR21","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1016\/S0014-2999(03)01274-3","volume":"463","author":"M Bourin","year":"2003","unstructured":"Bourin, M. & Hasco\u00ebt, M. The mouse light\/dark box test. Eur. J. Pharmacol. 463, 55\u201365 (2003).","journal-title":"Eur. J. Pharmacol."},{"key":"4286_CR22","doi-asserted-by":"publisher","first-page":"2289","DOI":"10.1038\/s41598-023-29052-7","volume":"13","author":"D Segelcke","year":"2023","unstructured":"Segelcke, D. et al. Experimenter familiarization is a crucial prerequisite for assessing behavioral outcomes and reducing stress in mice not only under chronic pain conditions. Sci. Rep. 13, 2289 (2023).","journal-title":"Sci. Rep."},{"key":"4286_CR23","doi-asserted-by":"publisher","first-page":"20305","DOI":"10.1038\/s41598-019-56860-7","volume":"9","author":"K Gouveia","year":"2019","unstructured":"Gouveia, K. & Hurst, J. L. Improving the practicality of using non-aversive handling methods to reduce background stress and anxiety in laboratory mice. Sci. Rep. 9, 20305 (2019).","journal-title":"Sci. Rep."},{"key":"4286_CR24","doi-asserted-by":"crossref","unstructured":"Riebe, C. J. & Wotjak, C. T. A Practical Guide to Evaluating Anxiety-Related Behavior in Rodents. In TRP Channels in Drug Discovery: Volume II (eds. Szallasi, A. & B\u00edr\u00f3, T.) 167\u2013185 (Humana Press, Totowa, NJ, 2012).","DOI":"10.1007\/978-1-62703-095-3_10"},{"key":"4286_CR25","doi-asserted-by":"publisher","first-page":"602","DOI":"10.4314\/jfas.v9i2s.38","volume":"9","author":"SB Mohamed","year":"2017","unstructured":"Mohamed, S. B. et al. An intelligent lighting control system (ILCS) using LabVIEW. J. Fundam. Appl. Sci. 9, 602\u2013615 (2017).","journal-title":"J. Fundam. Appl. Sci."},{"key":"4286_CR26","doi-asserted-by":"crossref","unstructured":"Ruddock, K. H. Light Transmission through the Ocular Media and Macular Pigment and its Significance for Psychophysical Investigation. In Visual Psychophysics (eds. Alpern, M. et al.) 455\u2013469 (Springer, Berlin, Heidelberg, 1972).","DOI":"10.1007\/978-3-642-88658-4_17"},{"key":"4286_CR27","doi-asserted-by":"publisher","first-page":"1221","DOI":"10.1038\/s41467-019-09124-x","volume":"10","author":"MH Berry","year":"2019","unstructured":"Berry, M. H. et al. Restoration of high-sensitivity and adapting vision with a cone opsin. Nat. Commun. 10, 1221 (2019).","journal-title":"Nat. Commun."},{"key":"4286_CR28","doi-asserted-by":"publisher","first-page":"1862","DOI":"10.1038\/s41467-017-01990-7","volume":"8","author":"MH Berry","year":"2017","unstructured":"Berry, M. H. et al. Restoration of patterned vision with an engineered photoactivatable G protein-coupled receptor. Nat. Commun. 8, 1862 (2017).","journal-title":"Nat. Commun."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-04286-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-04286-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-04286-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T16:39:42Z","timestamp":1751387982000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-025-04286-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,1]]},"references-count":28,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["4286"],"URL":"https:\/\/doi.org\/10.1038\/s41598-025-04286-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,1]]},"assertion":[{"value":"24 December 2024","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 May 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 July 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}},{"value":"The animal study was approved by the Animal Care and Use Committee (ACUC) at University of California, Berkeley. The study was conducted in accordance with the local legislation and institutional requirements.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics statement"}}],"article-number":"21204"}}