Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines.
Genetic medicines, including CRISPR/Cas technologies, extend tremendous promise for addressing unmet medical need in inherited retinal disorders and other indications; however, there remain challenges for the development of therapeutics. Herein, we evaluate genome editing by engineered Cas9 ribonucl...
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Public Library of Science (PLoS)
2025-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0317387 |
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| author | Spencer C Wei Aaron J Cantor Jack Walleshauser Rina Mepani Kory Melton Ashil Bans Prachi Khekare Suhani Gupta Jonathan Wang Craig Soares Radwan Kiwan Jieun Lee Shannon McCawley Vihasi Jani Weng In Leong Pawan K Shahi Jean Chan Pierre Boivin Peter Otoupal Bikash R Pattnaik David M Gamm Krishanu Saha Benjamin G Gowen Mary Haak-Frendscho Mary J Janatpour Adam P Silverman |
| author_facet | Spencer C Wei Aaron J Cantor Jack Walleshauser Rina Mepani Kory Melton Ashil Bans Prachi Khekare Suhani Gupta Jonathan Wang Craig Soares Radwan Kiwan Jieun Lee Shannon McCawley Vihasi Jani Weng In Leong Pawan K Shahi Jean Chan Pierre Boivin Peter Otoupal Bikash R Pattnaik David M Gamm Krishanu Saha Benjamin G Gowen Mary Haak-Frendscho Mary J Janatpour Adam P Silverman |
| author_sort | Spencer C Wei |
| collection | DOAJ |
| description | Genetic medicines, including CRISPR/Cas technologies, extend tremendous promise for addressing unmet medical need in inherited retinal disorders and other indications; however, there remain challenges for the development of therapeutics. Herein, we evaluate genome editing by engineered Cas9 ribonucleoproteins (eRNP) in vivo via subretinal administration using mouse and pig animal models. Subretinal administration of adenine base editor and double strand break-inducing Cas9 nuclease eRNPs mediate genome editing in both species. Editing occurs in retinal pigmented epithelium (RPE) and photoreceptor cells, with favorable tolerability in both species. Using transgenic reporter strains, we determine that editing primarily occurs close to the site of administration, within the bleb region associated with subretinal injection. Our results show that subretinal administration of BE-eRNPs in mice mediates base editing of up to 12% of the total neural retina, with an average rate of 7% observed at the highest dose tested. In contrast, a substantially lower editing efficiency was observed in minipigs; even with direct quantification of only the treated region, a maximum base editing rate of 1.5%, with an average rate of <1%, was observed. Our data highlight the importance of species consideration in preclinical studies for the development of genetic medicines targeting the eye and provide an example of a lack of translation between small and larger animal models in the context of subretinal administration of Cas9 eRNPs. |
| format | Article |
| id | doaj-art-8b17b71bf6384e7eb8b7c60e32f9518b |
| institution | Matheson Library |
| issn | 1932-6203 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Public Library of Science (PLoS) |
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| series | PLoS ONE |
| spelling | doaj-art-8b17b71bf6384e7eb8b7c60e32f9518b2025-07-10T05:31:50ZengPublic Library of Science (PLoS)PLoS ONE1932-62032025-01-01206e031738710.1371/journal.pone.0317387Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines.Spencer C WeiAaron J CantorJack WalleshauserRina MepaniKory MeltonAshil BansPrachi KhekareSuhani GuptaJonathan WangCraig SoaresRadwan KiwanJieun LeeShannon McCawleyVihasi JaniWeng In LeongPawan K ShahiJean ChanPierre BoivinPeter OtoupalBikash R PattnaikDavid M GammKrishanu SahaBenjamin G GowenMary Haak-FrendschoMary J JanatpourAdam P SilvermanGenetic medicines, including CRISPR/Cas technologies, extend tremendous promise for addressing unmet medical need in inherited retinal disorders and other indications; however, there remain challenges for the development of therapeutics. Herein, we evaluate genome editing by engineered Cas9 ribonucleoproteins (eRNP) in vivo via subretinal administration using mouse and pig animal models. Subretinal administration of adenine base editor and double strand break-inducing Cas9 nuclease eRNPs mediate genome editing in both species. Editing occurs in retinal pigmented epithelium (RPE) and photoreceptor cells, with favorable tolerability in both species. Using transgenic reporter strains, we determine that editing primarily occurs close to the site of administration, within the bleb region associated with subretinal injection. Our results show that subretinal administration of BE-eRNPs in mice mediates base editing of up to 12% of the total neural retina, with an average rate of 7% observed at the highest dose tested. In contrast, a substantially lower editing efficiency was observed in minipigs; even with direct quantification of only the treated region, a maximum base editing rate of 1.5%, with an average rate of <1%, was observed. Our data highlight the importance of species consideration in preclinical studies for the development of genetic medicines targeting the eye and provide an example of a lack of translation between small and larger animal models in the context of subretinal administration of Cas9 eRNPs.https://doi.org/10.1371/journal.pone.0317387 |
| spellingShingle | Spencer C Wei Aaron J Cantor Jack Walleshauser Rina Mepani Kory Melton Ashil Bans Prachi Khekare Suhani Gupta Jonathan Wang Craig Soares Radwan Kiwan Jieun Lee Shannon McCawley Vihasi Jani Weng In Leong Pawan K Shahi Jean Chan Pierre Boivin Peter Otoupal Bikash R Pattnaik David M Gamm Krishanu Saha Benjamin G Gowen Mary Haak-Frendscho Mary J Janatpour Adam P Silverman Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines. PLoS ONE |
| title | Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines. |
| title_full | Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines. |
| title_fullStr | Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines. |
| title_full_unstemmed | Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines. |
| title_short | Evaluation of subretinally delivered Cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines. |
| title_sort | evaluation of subretinally delivered cas9 ribonucleoproteins in murine and porcine animal models highlights key considerations for therapeutic translation of genetic medicines |
| url | https://doi.org/10.1371/journal.pone.0317387 |
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