The CASP6 Knockout HCT 116 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line. This polyclonal pool harbors a heterogeneous collection of CASP6 gene disruptions introduced by CRISPR/Cas9-mediated targeting, resulting in a loss-of-function model for the executioner caspase-6. Because the population is not clonal, it retains the genetic diversity of multiple editing events, making it suitable for studying bulk cellular responses without the bias of single-cell isolation. The knockout product format enables robust experimental designs that rely on pooled phenotypes, such as drug-screening campaigns or pathway interrogation in a cancer-relevant background.
The host cell line, HCT 116, is an adherent epithelial line originally isolated from a human colorectal carcinoma. These cells are widely employed in oncology research due to their well-characterized mutational landscape, stable karyotype, and responsiveness to apoptotic stimuli. HCT 116 cells express key components of both the extrinsic and intrinsic apoptotic machinery, making them an ideal platform for dissecting caspase-dependent cell death mechanisms. Their use in functional genomics, particularly in the context of colorectal cancer, provides a physiologically relevant system for evaluating tumor cell apoptosis and therapeutic vulnerabilities.
CASP6 encodes caspase-6, a cysteine-aspartic protease that functions as a downstream executioner in the apoptotic cascade. Proteolytic activation of pro-caspase-6 is carried out by initiator caspases such as caspase-8 and caspase-9, as well as by effector caspases caspase-3 and caspase-7, and the cytotoxic lymphocyte-derived serine protease granzyme B. Once activated, caspase-6 cleaves specific substrates including lamin A/C, cytokeratin 18, and the chromatin organizer SATB1, leading to nuclear lamina disassembly and cellular dismantling. Caspase-6 operates within a network that includes upstream death receptors (FAS), adaptor proteins (FADD), BCL-2 family members (BID, BAX, BAK), cytochrome c, APAF-1, and the apoptosome, thereby integrating signals from both extrinsic and intrinsic apoptosis pathways.
Disruption of CASP6 in HCT 116 cells provides a powerful tool to investigate the contribution of this executioner caspase to colorectal cancer cell apoptosis. The knockout model allows researchers to dissect the specific roles of caspase-6 downstream of death receptor ligation or chemotherapeutic stress, and to identify caspase-6-selective substrates whose cleavage may be critical for tumor cell death. Because HCT 116 cells are proficient in apoptosis, loss of CASP6 can reveal compensatory or alternative cell death pathways, offering insights into resistance mechanisms that limit the efficacy of anticancer agents. This model is particularly relevant for studying the execution phase of apoptosis in a cancer cell context, where caspase-6 activity has been implicated in the response to DNA-damaging drugs and targeted therapies.
The CASP6 Knockout HCT 116 Polyclonal Cells are suited for a range of advanced research applications, including the validation of caspase-6 substrates by western blot detection of cleaved lamin A/C or cytokeratin 18, measurement of caspase-6 enzymatic activity using fluorogenic substrates, and flow cytometric quantification of apoptosis induced by staurosporine or TRAIL using Annexin V/PI staining. Additional applications encompass cell viability assays (MTT) to assess chemosensitivity, immunofluorescence imaging of lamin A integrity, and RT-qPCR confirmation of CASP6 transcript loss. This polyclonal knockout population can also be employed in screens for small-molecule caspase-6 inhibitors or in genetic complementation studies. For further technical details or custom requests, please contact Ascent Research.
