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Elektroporationsbaseret genetisk modifikation af primære humane pigmentepitelceller ved hjælp af Sleeping Beauty Transposon-systemet
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Elektroporationsbaseret genetisk modifikation af primære humane pigmentepitelceller ved hjælp af Sleeping Beauty Transposon-systemet

Article DOI: 10.3791/61987-v • 07:04 min • February 4th, 2021
February 4th, 2021 •
1, 2,3, 4,5, 4, 6, 7, 1, 2,3
1Department of Ophthalmology, University Hospital RWTH Aachen, 2Experimental Ophthalmology, University of Geneva, 3Department of Ophthalmology, University Hospitals of Geneva, 4Université de Paris, CNRS, INSERM, UTCBS, Unité des technologies Chimiques et Biologiques pour la Santé, 5Chimie ParisTech, PSL Research University, 6Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 7Division of Medical Biotechnology, Paul-Ehrlich-Institute

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Vi har udviklet en protokol til at transficere primære humane pigmentepitelceller ved elektroporation med genet, der koder for pigmentepitelafledt faktor (PEDF) ved hjælp af Sleeping Beauty (SB) transposonsystemet. Vellykket transfektion blev påvist ved kvantitativ polymerasekædereaktion (qPCR), immunoblotting og enzymbundet immunosorbentassay (ELISA).

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Medicin udgave 168 aldersrelateret makuladegeneration AMD kapillær elektroporationssystem ikke-viral transfektion pigmentepitelafledt faktor PEDF primære pigmentepitelceller Tornerose transposon system SB
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