Generation of hZBP1-expressing Lentiviral Vectors

To generate human ZBP1 (hZBP1)-expressing lentiviral vector, 3XFLAG epitope-tagged hZBP1 open reading was inserted into retroviral expression vector pQCXIH (Clontech). Murine ZBP1 (mZBP1) and mutants inserted in pQCXIH were described previously^{22 (link)}. All plasmids were verified by DNA sequencing. Retrovirus stock preparation and transduction were described previously^{14 (link)}. Reconstituted HT-29 or SVEC4-10 cell lines were selected in 500 or 400 μg/ml hygromycin, respectively.

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Guo H., Gilley R.P., Fisher A., Lane R., Landsteiner V.J., Ragan K.B., Dovey C.M., Carette J.E., Upton J.W., Mocarski E.S, & Kaiser W.J. (2018). Species-independent contribution of ZBP1/DAI/DLM-1-triggered necroptosis in host defense against HSV1. Cell Death & Disease, 9(8), 816.

Publication 2018

pQCXIH

Cell lines Epitope Ht 29 Human Hygromycin Murine Plasmids Retrovirus Vector

Corresponding Organization :

Other organizations : The University of Texas at San Antonio, Stanford University, Emory University

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Variable analysis

independent variables

HZBP1 expression vector (pQCXIH with 3XFLAG epitope-tagged hZBP1 open reading frame)
MZBP1 and mutants expressed in pQCXIH

dependent variables

Reconstituted HT-29 or SVEC4-10 cell lines

control variables

Hygromycin selection concentration (500 μg/ml for HT-29, 400 μg/ml for SVEC4-10)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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