Matches in Ghent University Academic Bibliography for { <https://biblio.ugent.be/publication/01HMRQEWWGHQRV3XMMSMWB5ZEY> ?p ?o. }
Showing items 1 to 23 of
23
with 100 items per page.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY classification C3.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY date "2022".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY language "eng".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY type conference.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY hasPart 01HMRQNXD9QP106EW85D639P3W.pdf.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY subject "Biology and Life Sciences".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY subject "Medicine and Health Sciences".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY presentedAt urn:uuid:c9aedb92-8b25-4fb0-b282-954aa106609c.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY abstract "Background Inherited retinal diseases (IRDs) are a clinically and genetically heterogenous group of disorders leading to early-onset vision loss. RPE65 is a known IRD disease gene, encoding a crucial enzyme of the visual cycle that is important for preserving vision. In 2017-2018, Luxturna received FDA and EMA approval, respectively, as first gene therapy to treat IRD patients with biallelic mutations in RPE65. Therapeutic eligibility however requires a clear-cut molecular diagnosis, which is often hampered by the identification of Variants of Uncertain Significance (VUS), providing insufficient evidence concerning their role in the disease and their effect on gene/protein function. This study aimed to develop an in vitro biochemical assay to assess the pathogenicity of coding RPE65 VUS. Methods and results The assay is based on simulating the visual cycle via overexpression of visual cycle enzymes including RPE65 in HEK293-F cells. As RPE65 is responsible for the isomerization of all-trans retinyl esters to 11-cis retinol, the read-out consists of HPLC-based analysis of residual retinols, as well as immunoblotting of RPE65 protein. First, missense variants that had not been assessed functionally were compiled from various sources including our in-house database, LOVD, and members of the European Retinal Disease Consortium (ERDC) and the European Reference Network for Rare Eye Diseases (ERN-EYE). Variant classification according to ACMG/AMP guidelines revealed 53 RPE65 VUS to be assayed. Constructs have been generated by cloning the open reading frames of RPE65, CRALBP, LRAT and RDH5 in the multiple cloning sites (MCSs) of a pVitro2 backbone. Expression of bicistronic constructs has been compared to monocistronic constructs. Mutagenesis for the selected VUS has been performed and a first test assaying nine VUS and control variants has been completed. Conclusion Functional assessment of 53 RPE65 VUS via the enzymatic assay designed here will improve IRD patient eligibility for Luxturna gene therapy.".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY author 09227D4E-F0EE-11E1-A9DE-61C894A0A6B4.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY author 464708AE-3DED-11E4-8AC5-DAE6B4D1D7B1.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY author F50FFF8E-F0ED-11E1-A9DE-61C894A0A6B4.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY author FE51C442-F0ED-11E1-A9DE-61C894A0A6B4.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY author urn:uuid:6e99f334-ab49-4e8e-ba07-563a5529a88b.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY dateCreated "2024-01-22T13:58:48Z".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY dateModified "2024-10-29T18:36:50Z".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY name "An in vitro enzymatic assay to elucidate the VUS problem in RPE65, a target for retinal gene therapy".
- 01HMRQEWWGHQRV3XMMSMWB5ZEY pagination urn:uuid:498e8fe9-43a3-4848-98e5-9cae15f6c1d4.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY sameAs LU-01HMRQEWWGHQRV3XMMSMWB5ZEY.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY sourceOrganization urn:uuid:4561f271-9cf1-47f8-bba5-09ae48019131.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY sourceOrganization urn:uuid:4a9ce33d-623b-4d1c-8ad1-8aa4b2d99d21.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY sourceOrganization urn:uuid:87c1a64c-b698-40da-8758-b5031a0b6f0b.
- 01HMRQEWWGHQRV3XMMSMWB5ZEY type C3.