Lentiviral Vectors for Viroporin and mCAT Expression

The lentiviral vector used to co-express 2-E+2-3a was LV-E3a (Vectorbuilder ID VB210112-1153ufz) and its respective control vector LV-EV (Vectorbuilder ID VB210112-1153ufz). The LV-E3a vector contains the cytomegalovirus (CMV) promoter, the 2-E+2-3a viroporins obtained from Gordon et al., 2020 (18 (link)) separated by a 2A peptide site, and terminated by a SV40 late polyA sequence cloned into the LV-EV vector. The viroporin sequences deduced from the protein sequences were modified by adding an ATG codon 5’ and three N-terminal FLAG-tags added to the 3’ end of each viral protein. LV-EV is an empty vector.
The lentiviral vector expressing our mCAT and its respective control vector were LV-mCAT (VectorBuilder 210909-1242kdf) and LV-EV (VectorBuilder 900122-0484ubz). The LV-mCAT vector includes the EFS promoter, EGFP, 2A peptide site, mCAT, and SV40 late polyA sequence. The LV-EVcontrol vector lacks the EGFP and mCAT sequences.

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Guarnieri J.W., Angelin A., Murdock D.G., Schaefer P., Portluri P., Lie T., Huang J, & Wallace D.C. (2023). SARS-COV-2 viroporins activate the NLRP3-inflammasome by the mitochondrial permeability transition pore. Frontiers in Immunology, 14, 1064293.

Publication 2023

Codon Cytomegalovirus Peptide Polya Protein sequences Sv40 Vector Viral protein Viroporin

Corresponding Organization : University of Pennsylvania

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Variable analysis

independent variables

Presence of the LV-E3a vector (containing the 2-E+2-3a viroporin sequences) or the control LV-EV vector
Presence of the LV-mCAT vector (containing the mCAT sequence) or the control LV-EV vector

dependent variables

Outcomes or effects resulting from the expression of the 2-E+2-3a viroporin proteins or the mCAT protein

control variables

The LV-EV vector, which lacks the 2-E+2-3a viroporins or the mCAT sequence, serves as a control for the respective experimental vectors

controls

Positive control: LV-E3a vector (containing the 2-E+2-3a viroporin sequences)
Negative control: LV-EV vector (empty vector)

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