2020
DOI: 10.3390/ijms21030994
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AAV-Mediated Gene Delivery to 3D Retinal Organoids Derived from Human Induced Pluripotent Stem Cells

Abstract: Human induced pluripotent stem cells (hiPSCs) promise a great number of future applications to investigate retinal development, pathophysiology and cell therapies for retinal degenerative diseases. Specific approaches to genetically modulate hiPSC would be valuable for all of these applications. Vectors based on adeno-associated virus (AAV) have shown the ability for gene delivery to retinal organoids derived from hiPSCs. Thus far, little work has been carried out to investigate mechanisms of AAV-mediated gene… Show more

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Cited by 57 publications
(61 citation statements)
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“…AAV is a highly efficient method of delivering therapeutic genes to photoreceptor cells and AAV2/5 has been shown to effectively transduce photoreceptors in a variety of species inter alia mice ( Palfi et al., 2012 ), non-human primates ( Boye et al., 2012 ), and human retinal explants ( Wiley et al., 2018 ). The use of AAV in human ROs is currently limited, but studies have reported varying efficiency of transduction, which could be attributed to age of treatment, time of harvesting, vector tropisms, viral titer, the promotor used, and the sensitivity of detection of the transgene, e.g., antibody versus intrinsic fluorescence of a reporter ( Garita-Hernandez et al., 2020 , Gonzalez-Cordero et al., 2018 , Quinn et al., 2019 ). Here, we demonstrate highly efficient transduction using an AAV2/5 vector with RP2 under the control of a CAG promoter.…”
Section: Discussionmentioning
confidence: 99%
“…AAV is a highly efficient method of delivering therapeutic genes to photoreceptor cells and AAV2/5 has been shown to effectively transduce photoreceptors in a variety of species inter alia mice ( Palfi et al., 2012 ), non-human primates ( Boye et al., 2012 ), and human retinal explants ( Wiley et al., 2018 ). The use of AAV in human ROs is currently limited, but studies have reported varying efficiency of transduction, which could be attributed to age of treatment, time of harvesting, vector tropisms, viral titer, the promotor used, and the sensitivity of detection of the transgene, e.g., antibody versus intrinsic fluorescence of a reporter ( Garita-Hernandez et al., 2020 , Gonzalez-Cordero et al., 2018 , Quinn et al., 2019 ). Here, we demonstrate highly efficient transduction using an AAV2/5 vector with RP2 under the control of a CAG promoter.…”
Section: Discussionmentioning
confidence: 99%
“…We and other researchers use human retinal organoids to study rAAV transduction and potency [ 62 , 265 , 283 , 284 ]. In summary, photoreceptors are transduced by rAAVs such as rAAV2, rAAV2-7m8, rAAV5, rShH10, rShH10-Y445F, rAAV8, rAAV8T(Y733F), and rAAV9 albeit at different transduction efficacies.…”
Section: Transgene and Bioactivity Assays In Ocular Tissuementioning
confidence: 99%
“…The rAAV2-7m8, rAAV5, rShH10, and rShH10Y-445F capsids infect photoreceptors efficiently. Interestingly though, (early) radial retinal progenitor cells in retinal organoids or common cell lines can efficiently be infected by rAAV6, the rAAV6 variants (ShH10 and ShH10Y-445F), and the AAV2-7m8 [ 21 , 62 , 265 , 284 ]. For example, rAAV6 and rAAV-derived vectors (ShH10; ShH10-Y445F) infected hiPSCs and hiPSC-derived RPE cells properly and thoroughly [ 21 , 62 , 265 ].…”
Section: Transgene and Bioactivity Assays In Ocular Tissuementioning
confidence: 99%
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“…Since hydrogel matrices can often pose diffusional limitations to highly charged, relatively large non-viral gene delivery vectors, limitations remain in using CRISPR Cas9 to study organoids. Nevertheless, viral vectors are being employed together with nucleofection and electroporation to deliver genes to spheroid and organoid cultures ( Matano et al, 2015 ; Bian et al, 2018 ; Garita-Hernandez et al, 2020 ). The development of novel gene delivery tools can expand our ability to carry out direct genomic editing of organoids.…”
Section: Crispr Cas9 Systemsmentioning
confidence: 99%