{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T14:27:39Z","timestamp":1780496859708,"version":"3.54.1"},"publisher-location":"New York, NY","reference-count":14,"publisher":"Springer US","isbn-type":[{"value":"9781071620090","type":"print"},{"value":"9781071620106","type":"electronic"}],"license":[{"start":{"date-parts":[[2022,1,1]],"date-time":"2022-01-01T00:00:00Z","timestamp":1640995200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,1,1]],"date-time":"2022-01-01T00:00:00Z","timestamp":1640995200000},"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":[],"published-print":{"date-parts":[[2022]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Inherited retinal dystrophies, such as Leber congenital amaurosis, Stargardt disease, and retinitis pigmentosa, are characterized by photoreceptor dysfunction and death and currently have few treatment options. Recent technological advances in induced pluripotent stem cell (iPSC) technology and differentiation methods mean that human photoreceptors can now be studied in vitro. For example, retinal organoids provide a platform to study the development of the human retina and mechanisms of diseases in the dish, as well as being a potential source for cell transplantation. Here, we describe differentiation protocols for 3D cultures that produce retinal organoids containing photoreceptors with rudimentary outer segments. These protocols can be used as a model to understand retinal disease mechanisms and test potential therapies, including antisense oligonucleotides (AONs) to alter gene expression or RNA processing. This \u201cretina in a dish\u201d model is well suited for use with AONs, as the organoids recapitulate patient mutations in the correct genomic and cellular context, to test potential efficacy and examine off-target effects on the translational path to the clinic.<\/jats:p>","DOI":"10.1007\/978-1-0716-2010-6_16","type":"book-chapter","created":{"date-parts":[[2022,2,25]],"date-time":"2022-02-25T11:11:35Z","timestamp":1645787495000},"page":"245-255","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Eye on a Dish Models to Evaluate Splicing Modulation"],"prefix":"10.1007","author":[{"given":"Kwan-Leong","family":"Hau","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amelia","family":"Lane","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosellina","family":"Guarascio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael E.","family":"Cheetham","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2022,2,25]]},"reference":[{"issue":"8","key":"16_CR1","doi-asserted-by":"publisher","first-page":"741","DOI":"10.1038\/nbt.2643","volume":"31","author":"A Gonzalez-Cordero","year":"2013","unstructured":"Gonzalez-Cordero A, West EL, Pearson RA, Duran Y, Carvalho LS, Chu CJ, Naeem A, Blackford SJI, Georgiadis A, Lakowski J, Hubank M, Smith AJ, Bainbridge JWB, Sowden JC, Ali RR (2013) Photoreceptor precursors derived from three-dimensional embryonic stem cell cultures integrate and mature within adult degenerate retina. Nat Biotechnol 31(8):741\u2013747. https:\/\/doi.org\/10.1038\/nbt.2643","journal-title":"Nat Biotechnol"},{"key":"16_CR2","doi-asserted-by":"publisher","first-page":"4047","DOI":"10.1038\/ncomms5047","volume":"5","author":"X Zhong","year":"2014","unstructured":"Zhong X, Gutierrez C, Xue T, Hampton C, Vergara MN, Cao LH, Peters A, Park TS, Zambidis ET, Meyer JS, Gamm DM, Yau KW, Canto-Soler MV (2014) Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs. Nat Commun 5:4047. https:\/\/doi.org\/10.1038\/ncomms5047","journal-title":"Nat Commun"},{"issue":"6","key":"16_CR3","doi-asserted-by":"publisher","first-page":"771","DOI":"10.1016\/j.stem.2012.05.009","volume":"10","author":"T Nakano","year":"2012","unstructured":"Nakano T, Ando S, Takata N, Kawada M, Muguruma K, Sekiguchi K, Saito K, Yonemura S, Eiraku M, Sasai Y (2012) Self-formation of optic cups and storable stratified neural retina from human ESCs. Cell Stem Cell 10(6):771\u2013785. https:\/\/doi.org\/10.1016\/j.stem.2012.05.009","journal-title":"Cell Stem Cell"},{"issue":"39","key":"16_CR4","doi-asserted-by":"publisher","first-page":"16698","DOI":"10.1073\/pnas.0905245106","volume":"106","author":"JS Meyer","year":"2009","unstructured":"Meyer JS, Shearer RL, Capowski EE, Wright LS, Wallace KA, McMillan EL, Zhang SC, Gamm DM (2009) Modeling early retinal development with human embryonic and induced pluripotent stem cells. Proc Natl Acad Sci U S A 106(39):16698\u201316703. https:\/\/doi.org\/10.1073\/pnas.0905245106","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"3","key":"16_CR5","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1586\/eop.13.14","volume":"8","author":"DM Gamm","year":"2013","unstructured":"Gamm DM, Phillips MJ, Singh R (2013) Modeling retinal degenerative diseases with human iPS-derived cells: current status and future implications. Expert Rev Ophthalmol 8(3):213\u2013216. https:\/\/doi.org\/10.1586\/eop.13.14","journal-title":"Expert Rev Ophthalmol"},{"issue":"5","key":"16_CR6","doi-asserted-by":"publisher","first-page":"520","DOI":"10.1016\/j.stem.2013.04.009","volume":"12","author":"Y Sasai","year":"2013","unstructured":"Sasai Y (2013) Next-generation regenerative medicine: organogenesis from stem cells in 3D culture. Cell Stem Cell 12(5):520\u2013530. https:\/\/doi.org\/10.1016\/j.stem.2013.04.009","journal-title":"Cell Stem Cell"},{"issue":"9","key":"16_CR7","doi-asserted-by":"publisher","first-page":"565","DOI":"10.1016\/j.tins.2012.05.004","volume":"35","author":"EA Bassett","year":"2012","unstructured":"Bassett EA, Wallace VA (2012) Cell fate determination in the vertebrate retina. Trends Neurosci 35(9):565\u2013573. https:\/\/doi.org\/10.1016\/j.tins.2012.05.004","journal-title":"Trends Neurosci"},{"issue":"3","key":"16_CR8","doi-asserted-by":"publisher","first-page":"820","DOI":"10.1016\/j.stemcr.2017.07.022","volume":"9","author":"A Gonzalez-Cordero","year":"2017","unstructured":"Gonzalez-Cordero A, Kruczek K, Naeem A, Fernando M, Kloc M, Ribeiro J, Goh D, Duran Y, Blackford SJI, Abelleira-Hervas L, Sampson RD, Shum IO, Branch MJ, Gardner PJ, Sowden JC, Bainbridge JWB, Smith AJ, West EL, Pearson RA, Ali RR (2017) Recapitulation of human retinal development from human pluripotent stem cells generates transplantable populations of cone photoreceptors. Stem Cell Rep 9(3):820\u2013837. https:\/\/doi.org\/10.1016\/j.stemcr.2017.07.022","journal-title":"Stem Cell Rep"},{"issue":"6","key":"16_CR9","doi-asserted-by":"publisher","first-page":"769","DOI":"10.1016\/j.stem.2016.03.021","volume":"18","author":"DA Parfitt","year":"2016","unstructured":"Parfitt DA, Lane A, Ramsden CM, Carr AJ, Munro PM, Jovanovic K, Schwarz N, Kanuga N, Muthiah MN, Hull S, Gallo JM, da Cruz L, Moore AT, Hardcastle AJ, Coffey PJ, Cheetham ME (2016) Identification and correction of mechanisms underlying inherited blindness in human iPSC-derived optic cups. Cell Stem Cell 18(6):769\u2013781. https:\/\/doi.org\/10.1016\/j.stem.2016.03.021","journal-title":"Cell Stem Cell"},{"issue":"1","key":"16_CR10","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1038\/s41467-019-14020-5","volume":"11","author":"JP Ling","year":"2020","unstructured":"Ling JP, Wilks C, Charles R, Leavey PJ, Ghosh D, Jiang L, Santiago CP, Pang B, Venkataraman A, Clark BS, Nellore A, Langmead B, Blackshaw S (2020) ASCOT identifies key regulators of neuronal subtype-specific splicing. Nat Commun 11(1):137. https:\/\/doi.org\/10.1038\/s41467-019-14020-5","journal-title":"Nat Commun"},{"issue":"22","key":"16_CR11","doi-asserted-by":"publisher","first-page":"10824","DOI":"10.1073\/pnas.1901572116","volume":"116","author":"S Kim","year":"2019","unstructured":"Kim S, Lowe A, Dharmat R, Lee S, Owen LA, Wang J, Shakoor A, Li Y, Morgan DJ, Hejazi AA, Cvekl A, DeAngelis MM, Zhou ZJ, Chen R, Liu W (2019) Generation, transcriptome profiling, and functional validation of cone-rich human retinal organoids. Proc Natl Acad Sci U S A 116(22):10824\u201310833. https:\/\/doi.org\/10.1073\/pnas.1901572116","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"1","key":"16_CR12","doi-asserted-by":"publisher","first-page":"4234","DOI":"10.1038\/s41467-018-06448-y","volume":"9","author":"A Buskin","year":"2018","unstructured":"Buskin A, Zhu L, Chichagova V, Basu B, Mozaffari-Jovin S, Dolan D, Droop A, Collin J, Bronstein R, Mehrotra S, Farkas M, Hilgen G, White K, Pan KT, Treumann A, Hallam D, Bialas K, Chung G, Mellough C, Ding Y, Krasnogor N, Przyborski S, Zwolinski S, Al-Aama J, Alharthi S, Xu Y, Wheway G, Szymanska K, McKibbin M, Inglehearn CF, Elliott DJ, Lindsay S, Ali RR, Steel DH, Armstrong L, Sernagor E, Urlaub H, Pierce E, Luhrmann R, Grellscheid SN, Johnson CA, Lako M (2018) Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa. Nat Commun 9(1):4234. https:\/\/doi.org\/10.1038\/s41467-018-06448-y","journal-title":"Nat Commun"},{"key":"16_CR13","doi-asserted-by":"publisher","first-page":"730","DOI":"10.1016\/j.omtn.2018.07.010","volume":"12","author":"K Dulla","year":"2018","unstructured":"Dulla K, Aguila M, Lane A, Jovanovic K, Parfitt DA, Schulkens I, Chan HL, Schmidt I, Beumer W, Vorthoren L, Collin RWJ, Garanto A, Duijkers L, Brugulat-Panes A, Semo M, Vugler AA, Biasutto P, Adamson P, Cheetham ME (2018) Splice-modulating oligonucleotide QR-110 restores CEP290 mRNA and function in human c.2991+1655A>G LCA10 models. Mol Ther Nucleic Acids 12:730\u2013740. https:\/\/doi.org\/10.1016\/j.omtn.2018.07.010","journal-title":"Mol Ther Nucleic Acids"},{"key":"16_CR14","doi-asserted-by":"publisher","unstructured":"Capowski EE, Samimi K, Mayerl SJ, Phillips MJ, Pinilla I, Howden SE, Saha J, Jansen AD, Edwards KL, Jager LD, Barlow K, Valiauga R, Erlichman Z, Hagstrom A, Sinha D, Sluch VM, Chamling X, Zack DJ, Skala MC, Gamm DM (2019) Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines. Development 146(1). https:\/\/doi.org\/10.1242\/dev.171686","DOI":"10.1242\/dev.171686"}],"container-title":["Methods in Molecular Biology","Antisense RNA Design, Delivery, and Analysis"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/978-1-0716-2010-6_16","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,22]],"date-time":"2022-04-22T00:32:55Z","timestamp":1650587575000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/978-1-0716-2010-6_16"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022]]},"ISBN":["9781071620090","9781071620106"],"references-count":14,"URL":"https:\/\/doi.org\/10.1007\/978-1-0716-2010-6_16","relation":{},"ISSN":["1064-3745","1940-6029"],"issn-type":[{"value":"1064-3745","type":"print"},{"value":"1940-6029","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022]]},"assertion":[{"value":"25 February 2022","order":1,"name":"first_online","label":"First Online","group":{"name":"ChapterHistory","label":"Chapter History"}}]}}