Molecular imaging of human embryonic stem cells stably expressing human PET reporter genes after zinc finger nucleases-mediated genome editing

Molecular imaging is indispensable for determining the fate and persistence of engrafted stem cells. Standard strategies for transgene induction involve the use of viral vectors prone to silencing and insertional mutagenesis or the use of non-human genes. We used zinc finger nucleases (ZFN) to induce stable expression of human imaging reporter genes into the safe harbor locus adeno-associated virus integration site 1 (AAVS1) in human embryonic stem cells (ESC). Plasmids were generated carrying reporter genes for fluorescence, bioluminescence imaging (BLI), and human positron emission tomography (PET) reporter genes. In vitro assays confirmed their functionality and ESC retained differentiation capacity. Teratoma formation assays were performed and tumors were imaged over time with PET and BLI. This study demonstrates the application of genome editing for targeted integration of human imaging reporter genes in human ESC for long-term molecular imaging.

Jaar van publicatie:2017

BOF-keylabel:ja

IOF-keylabel:ja

BOF-publication weight:10

CSS-citation score:1

Authors from:Higher Education

Toegankelijkheid:Closed