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• W4229332010 endingPage "679" @default.
• W4229332010 startingPage "679" @default.
• W4229332010 abstract "ABCA1 and ABCG1 are two ABC-transporters well-recognized to promote the efflux of cholesterol to apoAI and HDL, respectively. As these two ABC-transporters are critical to cholesterol metabolism, several studies have assessed the impact of ABCA1 and ABCG1 expression on cellular cholesterol homeostasis through ABC-transporter ablation or overexpressing ABCA1/ABCG1. However, for the latter, there are currently no well-established in vitro models to effectively induce long-term ABC-transporter expression in a variety of cultured cells. Therefore, we performed proof-of-principle in vitro studies to determine whether a LoxP-Stop-LoxP (LSL) system would provide Cre-inducible ABC-transporter expression. In our studies, we transfected HEK293 cells and the HEK293-derived cell line 293-Cre cells with ABCA1-LSL and ABCG1-LSL-based plasmids. Our results showed that while the ABCA1/ABCG1 protein expression was absent in the transfected HEK293 cells, the ABCA1 and ABCG1 protein expression was detected in the 293-Cre cells transfected with ABCA1-LSL and ABCG1-LSL, respectively. When we measured cholesterol efflux in transfected 293-Cre cells, we observed an enhanced apoAI-mediated cholesterol efflux in 293-Cre cells overexpressing ABCA1, and an HDL2-mediated cholesterol efflux in 293-Cre cells constitutively expressing ABCG1. We also observed an appreciable increase in HDL3-mediated cholesterol efflux in ABCA1-overexpressing 293-Cre cells, which suggests that ABCA1 is capable of effluxing cholesterol to small HDL particles. Our proof-of-concept experiments demonstrate that the LSL-system can be used to effectively regulate ABC-transporter expression in vitro, which, in turn, allows ABCA1/ABCG1-overexpression to be extensively studied at the cellular level." @default.
• W4229332010 created "2022-05-09" @default.
• W4229332010 creator A5025752362 @default.
• W4229332010 creator A5040573003 @default.
• W4229332010 creator A5048940806 @default.
• W4229332010 creator A5060052348 @default.
• W4229332010 date "2022-05-08" @default.
• W4229332010 modified "2023-10-14" @default.
• W4229332010 title "Utilizing the LoxP-Stop-LoxP System to Control Transgenic ABC-Transporter Expression In Vitro" @default.
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• W4229332010 doi "https://doi.org/10.3390/biom12050679" @default.
• W4229332010 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35625607" @default.
• W4229332010 hasPublicationYear "2022" @default.
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