The CASP7 Knockout HEK293T Polyclonal Cells from Ascent Research consist of a CRISPR/Cas9-edited heterogeneous polyclonal population of HEK293T cells engineered to disrupt the endogenous CASP7 locus. This knockout pool offers a loss-of-function model for studying caspase-7 biology without the need for single-cell cloning, allowing researchers to assess gene function in a cellular context that maintains the robust growth and transfectability properties of the parental line.
The HEK293T host cell line is a derivative of the original HEK293 human embryonic kidney epithelial cells that stably expresses the SV40 large T antigen. This modification enhances episomal replication of transfected plasmids containing the SV40 origin of replication, yielding high-level transient protein expression and efficient production of recombinant proteins and lentiviral or retroviral vectors. The 293T line is therefore a foundational tool in molecular biology, drug discovery, and virology.
CASP7 encodes caspase-7, an executioner caspase activated by initiator caspases, notably caspase-8 and caspase-9, downstream of death receptor and mitochondrial apoptotic signals. Upon proteolytic cleavage, active caspase-7 cleaves key substrates including PARP1, DFFA/ICAD, lamin A/C, ROCK1, and ??-fodrin, leading to DNA fragmentation, nuclear disassembly, and membrane blebbing. The pathway is regulated by inhibitors such as XIAP, which is counteracted by SMAC/DIABLO and HtrA2/Omi released from mitochondria. Thus, CASP7 functions as a critical effector in both intrinsic and extrinsic apoptotic cascades, propagating the cell death signal through extensive proteolysis.
Disruption of CASP7 in HEK293T cells yields a loss-of-function model that enables dissection of the executioner phase of apoptosis without confounding compensation from a closely related caspase-3. The 293T background is highly amenable to transient and stable gene delivery, allowing reconstitution with mutant or tagged caspase-7 for structure-function studies. Combined with its robust protein expression capabilities, this model facilitates the screening of chemical probes that selectively target caspase-7 over caspase-3 in a human cellular context.
This knockout product supports diverse apoptosis research, including cancer drug screening for compounds that overcome caspase-7 deficiency, neurodegenerative disease modeling, and high-throughput screening of caspase inhibitors or activators. Common readouts include cleaved caspase-7 and PARP1 immunoblotting, caspase-7 activity fluorometric assays, Annexin V/PI flow cytometry, and immunofluorescence for active caspase-7. The polyclonal population provides a cost-effective and reproducible reagent for routine assays. For further technical inquiries or custom bulk orders, please contact Ascent Research.
