CASP3 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T-lymphocyte line. This product features disruption of the CASP3 gene through CRISPR/Cas9-mediated gene disruption, generating a heterogeneous pool of cells with CASP3 loss-of-function. As a polyclonal population, it is well-suited for studying caspase-3-dependent apoptosis without the selection bias of clonal lines.
Jurkat cells are a CD4+ T-lymphocyte line isolated from an acute T cell leukemia patient, characterized by mutated p53 and PTEN deficiency. These genetic alterations render the cells sensitive to apoptotic stimuli and make them a classic model for investigating T cell signaling, immune response, and apoptosis. The well-defined signaling pathways in Jurkat cells enable detailed analysis of apoptotic mechanisms triggered by intrinsic and extrinsic cues.
Caspase-3 (CASP3) is an executioner caspase cleaved and activated by initiator caspases CASP8, CASP9, and CASP10 upon extrinsic death receptor (FAS, TNFRSF1A) or intrinsic mitochondrial (cytochrome c/APAF1) signals, as well as by granzyme B. Active caspase-3 cleaves PARP1, DFFA, ROCK1, and lamin A/C, leading to chromatin condensation, DNA fragmentation, and cell death. Inhibitors such as XIAP and BIRC5/survivin regulate its activity, and it functions within networks involving BCL2 family proteins, BID, and FADD. Thus, CASP3 knockout abrogates the final execution phase of apoptosis.
In Jurkat cells, CASP3 knockout impairs apoptosis execution induced by intrinsic and extrinsic stimuli, including death receptor ligands and chemotherapeutics. Combined with the p53 and PTEN defects, this creates a resistant model to dissect caspase-3-dependent and -independent cell death pathways such as necroptosis and autophagy. It enables study of upstream signaling events, BCL2 family regulation, and the role of cytochrome c release when executioner caspase function is compromised, providing insight into survival signaling crosstalk.
Applications for these polyclonal knockout cells include apoptosis resistance studies, drug screening for caspase-3-independent cytotoxic compounds, and validation of apoptosis pathway components in cancer, neurodegeneration, and autoimmunity. Compatible assays include Annexin V/PI flow cytometry, caspase activity measurements, Western blotting for cleaved caspase-3 and PARP, cell viability assays (MTT/XTT), DNA fragmentation analysis, and mitochondrial membrane potential assessment. These tools facilitate therapeutic target identification and mechanistic studies. For further information, please contact Ascent Research.
