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- W2912372546 abstract "Prions are infectious protein aggregates that cause several fatal neurodegenerative diseases. Prion research has been hindered by a lack of cellular paradigms for studying the replication of prions from different species. Although hamster prions have been widely used to study prion replication in animals and within in vitro amplification systems, they have proved challenging to propagate in cultured cells. Because the murine catecholaminergic cell line CAD5 is susceptible to a diverse range of mouse prion strains, we hypothesized that it might also be capable of propagating nonmouse prions. Here, using CRISPR/Cas9-mediated genome engineering, we demonstrate that CAD5 cells lacking endogenous mouse PrP expression (CAD5-PrP−/− cells) can be chronically infected with hamster prions following stable expression of hamster PrP. When exposed to the 263K, HY, or 139H hamster prion strains, these cells stably propagated high levels of protease-resistant PrP. Hamster prion replication required absence of mouse PrP, and hamster PrP inhibited the propagation of mouse prions. Cellular homogenates from 263K-infected cells exhibited prion seeding activity in the RT-QuIC assay and were infectious to naive cells expressing hamster PrP. Interestingly, murine N2a neuroblastoma cells ablated for endogenous PrP expression were susceptible to mouse prions, but not hamster prions upon expression of cognate PrP, suggesting that CAD5 cells either possess cellular factors that enhance or lack factors that restrict the diversity of prion strains that can be propagated. We conclude that transfected CAD5-PrP−/− cells may be a useful tool for assessing the biology of prion strains and dissecting the mechanism of prion replication." @default.
- W2912372546 created "2019-02-21" @default.
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- W2912372546 date "2019-03-01" @default.
- W2912372546 modified "2023-10-15" @default.
- W2912372546 title "Engineering a murine cell line for the stable propagation of hamster prions" @default.
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- W2912372546 doi "https://doi.org/10.1074/jbc.ra118.007135" @default.
- W2912372546 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6442044" @default.
- W2912372546 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/30705093" @default.
- W2912372546 hasPublicationYear "2019" @default.
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