Silencing FAF1 Gene Expression

For silencing of FAF1 gene expression, the pGIPZ lentiviral vector, which contains FAF1-specific shRNA sequences was purchased from Open Biosystems. (http://www.openbiosystems.com). Lentiviruses were produced as previously described [27 (link)]. In brief, HEK293T cells were transiently transfected with packaging plasmids (psPAX2 and pMD2.VSV-G) and pGIPZ containing non-specific shRNA or FAF1-specific shRNA sequences using Lipofectamine 2000. At 48–72 hr post-transfection, virus-containing media was collected and filtrated (0.45 μm filter, Millipore).

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Kim T.H., Lee H.C., Kim J.H., Hewawaduge C.Y., Chathuranga K., Chathuranga W.A., Ekanayaka P., Wijerathne H.M., Kim C.J., Kim E, & Lee J.S. (2019). Fas-associated factor 1 mediates NADPH oxidase-induced reactive oxygen species production and proinflammatory responses in macrophages against Listeria infection. PLoS Pathogens, 15(8), e1008004.

Publication 2019

pGIPZ lentiviral vector

Cells Lentiviruses Lipofectamine 2000 Plasmids Shrna Silencing gene Transfection Vector Virus

Corresponding Organization : Chungnam National University

Other organizations : Korea Research Institute of Bioscience and Biotechnology, Korea University of Science and Technology

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Variable analysis

independent variables

FAF1-specific shRNA sequences in pGIPZ lentiviral vector
Non-specific shRNA sequences in pGIPZ lentiviral vector

dependent variables

FAF1 gene expression

control variables

Packaging plasmids (psPAX2 and pMD2.VSV-G)
Lipofectamine 2000 transfection
Time points (48-72 hr post-transfection)
Virus-containing media collection and filtration (0.45 μm filter)

negative control

Non-specific shRNA sequences in pGIPZ lentiviral vector

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