Peer-reviewedCC BY-NC-NDSTAR protocols
Protocol for high-throughput identification of genes involved in pyroptotic cell death by live microscopy.
4 affiliations
- 1Laboratory of Intracellular Immunity, Institute of Biochemistry and Biophysics of the Polish Academy of Sciences, Warsaw, Poland; Doctoral School of Molecular Biology and Biological Chemistry, Institute of Biochemistry and Biophysics of the Polish Academy of Sciences, Warsaw, Poland. Electronic address: [email protected].
- 2Laboratory of RNA Biology, Institute of Biochemistry and Biophysics of the Polish Academy of Sciences, Warsaw, Poland; Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Warsaw, Poland.
- 3Laboratory of RNA Biology, Institute of Biochemistry and Biophysics of the Polish Academy of Sciences, Warsaw, Poland.
- 4Laboratory of Intracellular Immunity, Institute of Biochemistry and Biophysics of the Polish Academy of Sciences, Warsaw, Poland. Electronic address: [email protected].
Abstract
Intracellular lipopolysaccharide activates caspase-4/5 triggering pyroptotic cell death. We present a protocol for a high-throughput loss-of-function screening approach to identify interferon-gamma-stimulated genes regulating this pathway. We detail procedures for cell reverse-transfection with a custom-made small interfering RNA (siRNA) library in 384-well plates and priming with interferon-gamma. We then describe steps for transfection with lipopolysaccharide to induce pyroptosis and live-cell fluorescence microscopy using the IncuCyte S3 system to quantify cell death in real time without fixation. This approach enables reliable, high-throughput screening of cell death.
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BibTeX
@article{bajger2026protocol,
title = {{Protocol for high-throughput identification of genes involved in pyroptotic cell death by live microscopy.}},
author = {Aleksandra Bajger and Lukasz S Borowski and Roman J Szczesny and Michal P Wandel},
journal = {STAR protocols},
year = {2026},
month = oct,
doi = {10.1016/j.xpro.2026.104884},
url = {https://europepmc.org/article/MED/42848516},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Protocol for high-throughput identification of genes involved in pyroptotic cell death by live microscopy.
AU - Aleksandra Bajger
AU - Lukasz S Borowski
AU - Roman J Szczesny
AU - Michal P Wandel
PY - 2026
DA - 2026/10/07
JO - STAR protocols
DO - 10.1016/j.xpro.2026.104884
UR - https://europepmc.org/article/MED/42848516
AB - Intracellular lipopolysaccharide activates caspase-4/5 triggering pyroptotic cell death. We present a protocol for a high-throughput loss-of-function screening approach to identify interferon-gamma-stimulated genes regulating this pathway. We detail procedures for cell reverse-transfection with a custom-made small interfering RNA (siRNA) library in 384-well plates and priming with interferon-gamma. We then describe steps for transfection with lipopolysaccharide to induce pyroptosis and live-cell fluorescence microscopy using the IncuCyte S3 system to quantify cell death in real time without fixation. This approach enables reliable, high-throughput screening of cell death.
SN - 2666-1667
ER -