{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,28]],"date-time":"2026-10-28T07:25:48Z","timestamp":1793172348460,"version":"4.5.0"},"reference-count":52,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2018,9,19]],"date-time":"2018-09-19T00:00:00Z","timestamp":1537315200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2018,9,19]],"date-time":"2018-09-19T00:00:00Z","timestamp":1537315200000},"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><jats:p>Optogenetic technologies have been the subject of great excitement within the scientific community for their ability to demystify complex neurophysiological pathways in the central (CNS) and peripheral nervous systems (PNS). The excitement surrounding optogenetics has also extended to the clinic with a trial for ChR2 in the treatment of retinitis pigmentosa currently underway and additional trials anticipated for the near future. In this work, we identify the cause of loss-of-expression in response to transdermal illumination of an optogenetically active peroneal nerve following an anterior compartment (AC) injection of AAV6-hSyn-ChR2(H134R) with and without a fluorescent reporter. Using Sprague Dawley Rag2<jats:sup>\u2212\/\u2212<\/jats:sup> rats and appropriate controls, we discover optogenetic loss-of-expression is chiefly elicited by ChR2-mediated immunogenicity in the spinal cord, resulting in both CNS motor neuron death and ipsilateral muscle atrophy in both low and high Adeno-Associated Virus (AAV) dosages. We further employ pharmacological immunosuppression using a slow-release tacrolimus pellet to demonstrate sustained transdermal optogenetic expression up to 12 weeks. These results suggest that all dosages of AAV-mediated optogenetic expression within the PNS may be unsafe. Clinical optogenetics for both PNS and CNS applications should take extreme caution when employing opsins to treat disease and may require concurrent immunosuppression. Future work in optogenetics should focus on designing opsins with lesser immunogenicity.<\/jats:p>","DOI":"10.1038\/s41598-018-32075-0","type":"journal-article","created":{"date-parts":[[2018,9,13]],"date-time":"2018-09-13T14:38:17Z","timestamp":1536849497000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":73,"title":["Optogenetic Peripheral Nerve Immunogenicity"],"prefix":"10.1038","volume":"8","author":[{"given":"Benjamin E.","family":"Maimon","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maurizio","family":"Diaz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emilie C. M.","family":"Revol","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2158-6915","authenticated-orcid":false,"given":"Alexis M.","family":"Schneider","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ben","family":"Leaker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia E.","family":"Varela","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2508-1324","authenticated-orcid":false,"given":"Shriya","family":"Srinivasan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew B.","family":"Weber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hugh M.","family":"Herr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2018,9,19]]},"reference":[{"key":"32075_CR1","doi-asserted-by":"publisher","first-page":"337rv5","DOI":"10.1126\/scitranslmed.aad7577","volume":"8","author":"KL Montgomery","year":"2016","unstructured":"Montgomery, K. L. et al. Beyond the brain: Optogenetic control in the spinal cord and peripheral nervous system. Sci. Transl. Med. 8, 337rv5 (2016).","journal-title":"Sci. Transl. Med."},{"key":"32075_CR2","doi-asserted-by":"publisher","first-page":"338","DOI":"10.1038\/nmeth.2836","volume":"11","author":"NC Klapoetke","year":"2014","unstructured":"Klapoetke, N. C. et al. Independent optical excitation of distinct neural populations. Nat. Methods 11, 338\u201346 (2014).","journal-title":"Nat. Methods"},{"key":"32075_CR3","doi-asserted-by":"publisher","first-page":"485","DOI":"10.1038\/s41551-018-0255-5","volume":"2","author":"BE Maimon","year":"2018","unstructured":"Maimon, B. E., Sparks, K., Srinivasan, S., Zorzos, A. N. & Herr, H. M. Spectrally distinct channelrhodopsins for two-colour optogenetic peripheral nerve stimulation. Nat. Biomed. Eng. 2, 485\u2013496 (2018).","journal-title":"Nat. Biomed. Eng."},{"key":"32075_CR4","doi-asserted-by":"publisher","DOI":"10.1088\/1741-2552\/aa5e20","author":"B Maimon","year":"2017","unstructured":"Maimon, B. et al. Transdermal optogenetic peripheral nerve stimulation. J. Neural Eng. https:\/\/doi.org\/10.1088\/1741-2552\/aa5e20 (2017).","journal-title":"J. Neural Eng"},{"key":"32075_CR5","doi-asserted-by":"publisher","first-page":"e72691","DOI":"10.1371\/journal.pone.0072691","volume":"8","author":"C Towne","year":"2013","unstructured":"Towne, C., Montgomery, K. L., Iyer, S. M., Deisseroth, K. & Delp, S. L. Optogenetic control of targeted peripheral axons in freely moving animals. PLoS One 8, e72691 (2013).","journal-title":"PLoS One"},{"key":"32075_CR6","first-page":"8","volume":"7","author":"T Miyashita","year":"2013","unstructured":"Miyashita, T., Shao, Y. R., Chung, J., Pourzia, O. & Feldman, D. E. Long-term channelrhodopsin-2 (ChR2) expression can induce abnormal axonal morphology and targeting in cerebral cortex. Front. Neural Circuits 7, 8 (2013).","journal-title":"Front. Neural Circuits"},{"key":"32075_CR7","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1007\/s11068-008-9027-6","volume":"36","author":"V Gradinaru","year":"2008","unstructured":"Gradinaru, V., Thompson, K. R. & Deisseroth, K. eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications. Brain Cell Biol. 36, 129\u2013139 (2008).","journal-title":"Brain Cell Biol."},{"key":"32075_CR8","doi-asserted-by":"publisher","first-page":"141","DOI":"10.1007\/s11068-008-9034-7","volume":"36","author":"S Zhao","year":"2008","unstructured":"Zhao, S. et al. Improved expression of halorhodopsin for light-induced silencing of neuronal activity. Brain Cell Biol. 36, 141\u2013154 (2008).","journal-title":"Brain Cell Biol."},{"key":"32075_CR9","doi-asserted-by":"publisher","first-page":"1022","DOI":"10.1016\/j.bbrc.2008.02.153","volume":"369","author":"D Zimmerman","year":"2008","unstructured":"Zimmerman, D. et al. Effects on capacitance by overexpression of membrane proteins. Biochem. Biophys. Res. Commun. 369, 1022\u20131026 (2008).","journal-title":"Biochem. Biophys. Res. Commun."},{"issue":"52","key":"32075_CR10","doi-asserted-by":"publisher","first-page":"14231","DOI":"10.1523\/JNEUROSCI.3578-07.2007","volume":"27","author":"V. Gradinaru","year":"2007","unstructured":"Gradinaru, V. et al. Targeting and Readout Strategies for Fast Optical Neural Control In Vitro and In Vivo. J. Neurosci. 27 (2007).","journal-title":"Journal of Neuroscience"},{"key":"32075_CR11","doi-asserted-by":"publisher","first-page":"473","DOI":"10.1080\/10915810290169909","volume":"21","author":"RD Glickman","year":"2002","unstructured":"Glickman, R. D. Phototoxicity to the Retina: Mechanisms of Damage. Int. J. Toxicol. 21, 473\u2013490 (2002).","journal-title":"Int. J. Toxicol."},{"key":"32075_CR12","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-00829-x","volume":"7","author":"JH Stockley","year":"2017","unstructured":"Stockley, J. H. et al. Surpassing light-induced cell damage in vitro with novel cell culture media. Sci. Rep. 7, 849 (2017).","journal-title":"Sci. Rep."},{"key":"32075_CR13","first-page":"10581","volume":"5","author":"I Khan","year":"2015","unstructured":"Khan, I., Tang, E. & Arany, P. Molecular pathway of near-infrared laser phototoxicity involves ATF-4 orchestrated ER. stress. 5, 10581 (2015).","journal-title":"stress."},{"key":"32075_CR14","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1016\/j.sbi.2006.10.001","volume":"16","author":"SJ Remington","year":"2006","unstructured":"Remington, S. J. Fluorescent proteins: maturation, photochemistry and photophysics. Curr. Opin. Struct. Biol. 16, 714\u2013721 (2006).","journal-title":"Curr. Opin. Struct. Biol."},{"key":"32075_CR15","doi-asserted-by":"publisher","first-page":"314","DOI":"10.1016\/j.neuron.2013.11.011","volume":"81","author":"K Beppu","year":"2014","unstructured":"Beppu, K. et al. Optogenetic Countering of Glial Acidosis Suppresses Glial Glutamate Release and Ischemic Brain Damage. Neuron 81, 314\u2013320 (2014).","journal-title":"Neuron"},{"key":"32075_CR16","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2016.351","volume":"7","author":"M Perny","year":"2016","unstructured":"Perny, M., Muri, L., Dawson, H. & Kleinlogel, S. Chronic activation of the D156A point mutant of Channelrhodopsin-2 signals apoptotic cell death: the good and the bad. Cell Death Dis. 7, e2447 (2016).","journal-title":"Cell Death Dis."},{"key":"32075_CR17","doi-asserted-by":"publisher","DOI":"10.1038\/srep14306","volume":"5","author":"Y Kohro","year":"2015","unstructured":"Kohro, Y. et al. A new minimally-invasive method for microinjection into the mouse spinal dorsal horn. Sci. Rep. 5, 14306 (2015).","journal-title":"Sci. Rep."},{"issue":"20","key":"32075_CR18","doi-asserted-by":"publisher","first-page":"1887","DOI":"10.1056\/NEJMoa1414221","volume":"372","author":"James W.B. Bainbridge","year":"2015","unstructured":"Bainbridge, J. W. B. et al. Long-Term Effect of Gene Therapy on Leber\u2019s Congenital Amaurosis. https:\/\/doi.org\/10.1056\/NEJMoa1414221 (2015).","journal-title":"New England Journal of Medicine"},{"key":"32075_CR19","doi-asserted-by":"publisher","first-page":"23","DOI":"10.1182\/blood-2013-01-306647","volume":"122","author":"F Mingozzi","year":"2013","unstructured":"Mingozzi, F. et al. Immune responses to AAV vectors: overcoming barriers to successful gene therapy. Blood 122, 23\u201336 (2013).","journal-title":"Blood"},{"key":"32075_CR20","doi-asserted-by":"publisher","first-page":"266","DOI":"10.1038\/gt.2010.140","volume":"18","author":"E Sugano","year":"2011","unstructured":"Sugano, E. et al. Immune responses to adeno-associated virus type 2 encoding channelrhodopsin-2 in a genetically blind rat model for gene therapy. Gene Ther. 18, 266\u2013274 (2011).","journal-title":"Gene Ther."},{"key":"32075_CR21","doi-asserted-by":"publisher","first-page":"399","DOI":"10.1002\/rmv.1762","volume":"23","author":"S Hareendran","year":"2013","unstructured":"Hareendran, S. et al. Adeno-associated virus (AAV) vectors in gene therapy: immune challenges and strategies to circumvent them. Rev. Med. Virol. 23, 399\u2013413 (2013).","journal-title":"Rev. Med. Virol."},{"key":"32075_CR22","doi-asserted-by":"publisher","first-page":"553","DOI":"10.1007\/s12015-016-9670-8","volume":"12","author":"AM Ansari","year":"2016","unstructured":"Ansari, A. M. et al. Cellular GFP Toxicity and Immunogenicity: Potential Confounders in in Vivo Cell Tracking Experiments. Stem Cell Rev. 12, 553\u2013559 (2016).","journal-title":"Stem Cell Rev."},{"key":"32075_CR23","doi-asserted-by":"publisher","first-page":"1305","DOI":"10.1038\/sj.gt.3300951","volume":"6","author":"R Stripecke","year":"1999","unstructured":"Stripecke, R. et al. Immune response to green fluorescent protein: implications for gene therapy. Gene Ther. 6, 1305\u20131312 (1999).","journal-title":"Gene Ther."},{"key":"32075_CR24","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1155\/2008\/453590","volume":"2008","author":"RR Taghizadeh","year":"2008","unstructured":"Taghizadeh, R. R. & Sherley, J. L. CFP and YFP, but Not GFP, Provide Stable Fluorescent Marking of Rat Hepatic Adult Stem Cells. J. Biomed. Biotechnol. 2008, 1\u20139 (2008).","journal-title":"J. Biomed. Biotechnol."},{"key":"32075_CR25","doi-asserted-by":"publisher","first-page":"1220","DOI":"10.1038\/mt.2011.69","volume":"19","author":"MM Doroudchi","year":"2011","unstructured":"Doroudchi, M. M. et al. Virally delivered Channelrhodopsin-2 Safely and Effectively Restores Visual Function in Multiple Mouse Models of Blindness. Mol. Ther. 19, 1220\u20131229 (2011).","journal-title":"Mol. Ther."},{"key":"32075_CR26","doi-asserted-by":"publisher","first-page":"1073","DOI":"10.1038\/mt.2008.76","volume":"16","author":"C Zincarelli","year":"2008","unstructured":"Zincarelli, C., Soltys, S., Rengo, G. & Rabinowitz, J. E. Analysis of AAV Serotypes 1\u20139 Mediated Gene Expression and Tropism in Mice After Systemic Injection. Mol. Ther. 16, 1073\u20131080 (2008).","journal-title":"Mol. Ther."},{"key":"32075_CR27","doi-asserted-by":"publisher","first-page":"e1002281","DOI":"10.1371\/journal.ppat.1002281","volume":"7","author":"LP Ganesan","year":"2011","unstructured":"Ganesan, L. P. et al. Rapid and Efficient Clearance of Blood-borne Virus by Liver Sinusoidal Endothelium. PLoS Pathog. 7, e1002281 (2011).","journal-title":"PLoS Pathog."},{"key":"32075_CR28","doi-asserted-by":"publisher","first-page":"651","DOI":"10.1016\/j.neuron.2004.10.026","volume":"44","author":"DL Bishop","year":"2004","unstructured":"Bishop, D. L., Misgeld, T., Walsh, M. K., Gan, W.-B. & Lichtman, J. W. Axon Branch Removal at Developing Synapses by Axosome Shedding. Neuron 44, 651\u2013661 (2004).","journal-title":"Neuron"},{"key":"32075_CR29","doi-asserted-by":"publisher","first-page":"63","DOI":"10.1038\/356063a0","volume":"356","author":"TA Yednock","year":"1992","unstructured":"Yednock, T. A. et al. Prevention of experimental autoimmune encephalomyelitis by antibodies against \u03b14\u03b2l integrin. Nature 356, 63\u201366 (1992).","journal-title":"Nature"},{"key":"32075_CR30","doi-asserted-by":"publisher","first-page":"271","DOI":"10.1016\/0272-0590(87)90049-2","volume":"9","author":"MJ Derelanko","year":"1987","unstructured":"Derelanko, M. J. Determination of erythrocyte life span in F-344, Wistar, and Sprague-Dawley rats using a modification of the [3H]diisopropylfluorophosphate ([3H]DFP) method. Fundam. Appl. Toxicol. 9, 271\u20136 (1987).","journal-title":"Fundam. Appl. Toxicol."},{"key":"32075_CR31","doi-asserted-by":"publisher","first-page":"2635","DOI":"10.1093\/jn\/133.8.2635","volume":"133","author":"SM Grant","year":"2003","unstructured":"Grant, S. M., Wiesinger, J. A., Beard, J. L. & Cantorna, M. T. Iron-deficient mice fail to develop autoimmune encephalomyelitis. J. Nutr. 133, 2635\u20138 (2003).","journal-title":"J. Nutr."},{"key":"32075_CR32","doi-asserted-by":"publisher","first-page":"M59","DOI":"10.1093\/gerona\/54.2.M59","volume":"54","author":"T F\u00fcl\u00f6p","year":"1999","unstructured":"F\u00fcl\u00f6p, T. et al. Relationship between the response to influenza vaccination and the nutritional status in institutionalized elderly subjects. J. Gerontol. A. Biol. Sci. Med. Sci. 54, M59\u201364 (1999).","journal-title":"J. Gerontol. A. Biol. Sci. Med. Sci."},{"key":"32075_CR33","doi-asserted-by":"publisher","first-page":"552","DOI":"10.1016\/j.clinbiochem.2005.02.011","volume":"38","author":"SH Akbas","year":"2005","unstructured":"Akbas, S. H. et al. Effects of some hematological parameters on whole blood tacrolimus concentration measured by two immunoassay-based analytical methods. Clin. Biochem. 38, 552\u2013557 (2005).","journal-title":"Clin. Biochem."},{"key":"32075_CR34","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-13952-6","volume":"7","author":"S Park","year":"2017","unstructured":"Park, S. et al. Association between post-transplant red cell distribution width and prognosis of kidney transplant recipients. Sci. Rep. 7, 13755 (2017).","journal-title":"Sci. Rep."},{"key":"32075_CR35","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1002\/ana.22421","volume":"70","author":"V Bartanusz","year":"2011","unstructured":"Bartanusz, V., Jezova, D., Alajajian, B. & Digicaylioglu, M. The blood-spinal cord barrier: Morphology and Clinical Implications. Ann. Neurol. 70, 194\u2013206 (2011).","journal-title":"Ann. Neurol."},{"key":"32075_CR36","doi-asserted-by":"publisher","first-page":"199","DOI":"10.3389\/fmicb.2011.00199","volume":"2","author":"S Adriouch","year":"2011","unstructured":"Adriouch, S. et al. Improved Immunological Tolerance Following Combination Therapy with CTLA-4\/Ig and AAV-Mediated PD-L1\/2 Muscle Gene Transfer. Front. Microbiol. 2, 199 (2011).","journal-title":"Front. Microbiol."},{"key":"32075_CR37","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1016\/j.expneurol.2013.05.017","volume":"248","author":"Juraj Sevc","year":"2013","unstructured":"Sevc, J. et al. Effective long-term immunosuppression in rats by subcutaneously implanted sustained-release tacrolimus pellet: Effect on spinally grafted human neural precursor survival. Exp. Neurol. 248, (2013).","journal-title":"Experimental Neurology"},{"key":"32075_CR38","doi-asserted-by":"publisher","first-page":"3081","DOI":"10.1111\/ajt.13399","volume":"15","author":"IR Badell","year":"2015","unstructured":"Badell, I. R. et al. Pathogen Stimulation History Impacts Donor-Specific CD8+ T Cell Susceptibility to Costimulation\/Integrin Blockade-Based Therapy. Am. J. Transplant. 15, 3081\u20133094 (2015).","journal-title":"Am. J. Transplant."},{"key":"32075_CR39","doi-asserted-by":"publisher","first-page":"e22453","DOI":"10.1371\/journal.pone.0022453","volume":"6","author":"AC-H Chen","year":"2011","unstructured":"Chen, A. C.-H. et al. Low-Level Laser Therapy Activates NF-kB via Generation of Reactive Oxygen Species in Mouse Embryonic Fibroblasts. PLoS One 6, e22453 (2011).","journal-title":"PLoS One"},{"key":"32075_CR40","doi-asserted-by":"publisher","first-page":"22","DOI":"10.3389\/fnmol.2013.00022","volume":"6","author":"G Lignani","year":"2013","unstructured":"Lignani, G. et al. Long-term optical stimulation of channelrhodopsin-expressing neurons to study network plasticity. Front. Mol. Neurosci. 6, 22 (2013).","journal-title":"Front. Mol. Neurosci."},{"key":"32075_CR41","doi-asserted-by":"publisher","first-page":"12317","DOI":"10.1073\/pnas.0905852106","volume":"106","author":"K Feldbauer","year":"2009","unstructured":"Feldbauer, K. et al. Channelrhodopsin-2 is a leaky proton pump. Proc. Natl. Acad. Sci. USA 106, 12317\u201322 (2009).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"32075_CR42","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-01723-2","volume":"7","author":"Q Li","year":"2017","unstructured":"Li, Q. et al. Electrophysiological Properties and Viability of Neonatal Rat Ventricular Myocyte Cultures with Inducible ChR2 Expression. Sci. Rep. 7, 1531 (2017).","journal-title":"Sci. Rep."},{"key":"32075_CR43","first-page":"493","volume":"11","author":"BK Sack","year":"2009","unstructured":"Sack, B. K. & Herzog, R. W. Evading the immune response upon in vivo gene therapy with viral vectors. Curr. Opin. Mol. Ther. 11, 493\u2013503 (2009).","journal-title":"Curr. Opin. Mol. Ther."},{"key":"32075_CR44","doi-asserted-by":"publisher","first-page":"819","DOI":"10.1146\/annurev.genet.37.110801.143717","volume":"38","author":"DM McCarty","year":"2004","unstructured":"McCarty, D. M., Young, S. M. & Samulski, R. J. Integration of Adeno-Associated Virus (AAV) and Recombinant AAV Vectors. Annu. Rev. Genet. 38, 819\u2013845 (2004).","journal-title":"Annu. Rev. Genet."},{"key":"32075_CR45","doi-asserted-by":"publisher","first-page":"306","DOI":"10.1038\/nm0397-306","volume":"3","author":"KJ Fisher","year":"1997","unstructured":"Fisher, K. J. et al. Recombinant adeno-associated virus for muscle directed gene therapy. Nat. Med. 3, 306\u201312 (1997).","journal-title":"Nat. Med."},{"key":"32075_CR46","doi-asserted-by":"publisher","first-page":"1465","DOI":"10.1093\/nar\/24.8.1465","volume":"24","author":"G Robertson","year":"1996","unstructured":"Robertson, G., Garrick, D., Wilson, M., Martin, D. I. & Whitelaw, E. Age-dependent silencing of globin transgenes in the mouse. Nucleic Acids Res. 24, 1465\u201371 (1996).","journal-title":"Nucleic Acids Res."},{"key":"32075_CR47","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1016\/j.ymthe.2005.11.010","volume":"13","author":"T Okada","year":"2006","unstructured":"Okada, T. et al. A Histone Deacetylase Inhibitor Enhances Recombinant Adeno-associated Virus-Mediated Gene Expression in Tumor Cells. Mol. Ther. 13, 738\u2013746 (2006).","journal-title":"Mol. Ther."},{"key":"32075_CR48","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1016\/S0378-1119(00)00353-X","volume":"256","author":"AR Migliaccio","year":"2000","unstructured":"Migliaccio, A. R. et al. Stable and unstable transgene integration sites in the human genome: extinction of the Green Fluorescent Protein transgene in K562 cells. Gene 256, 197\u2013214 (2000).","journal-title":"Gene"},{"issue":"4","key":"32075_CR49","doi-asserted-by":"publisher","first-page":"600","DOI":"10.1016\/j.ymthe.2004.10.020","volume":"11","author":"Kristi Jennings","year":"2005","unstructured":"Jennings, K. et al. Proteasome Inhibition Enhances AAV-Mediated Transgene Expression in Human Synoviocytes in Vitro and in Vivo. Mol. Ther. 11, 600\u2013607 (2005).","journal-title":"Molecular Therapy"},{"key":"32075_CR50","doi-asserted-by":"publisher","first-page":"e6679","DOI":"10.1371\/journal.pone.0006679","volume":"4","author":"Q Kong","year":"2009","unstructured":"Kong, Q. et al. Transgene Expression Is Associated with Copy Number and Cytomegalovirus Promoter Methylation in Transgenic Pigs. PLoS One 4, e6679 (2009).","journal-title":"PLoS One"},{"key":"32075_CR51","doi-asserted-by":"publisher","first-page":"R37","DOI":"10.1088\/0031-9155\/58\/11\/R37","volume":"58","author":"SL Jacques","year":"2013","unstructured":"Jacques, S. L. Optical properties of biological tissues: a review. Phys. Med. Biol. 58, R37\u201361 (2013).","journal-title":"Phys. Med. Biol."},{"key":"32075_CR52","doi-asserted-by":"publisher","first-page":"663","DOI":"10.3389\/fnins.2017.00663","volume":"11","author":"J Delbeke","year":"2017","unstructured":"Delbeke, J., Hoffman, L., Mols, K., Braeken, D. & Prodanov, D. And Then There Was Light: Perspectives of Optogenetics for Deep Brain Stimulation and Neuromodulation. Front. Neurosci. 11, 663 (2017).","journal-title":"Front. Neurosci."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-32075-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-32075-0","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-32075-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,21]],"date-time":"2022-12-21T09:39:29Z","timestamp":1671615569000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-018-32075-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,9,19]]},"references-count":52,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["32075"],"URL":"https:\/\/doi.org\/10.1038\/s41598-018-32075-0","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,9,19]]},"assertion":[{"value":"15 May 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 August 2018","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 September 2018","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"14076"}}