The CARD19 Knockout HAP1 Polyclonal Cells product comprises a heterogeneous population of HAP1 cells engineered via CRISPR/Cas9-mediated disruption of the CARD19 gene. This polyclonal knockout pool provides a versatile loss-of-function model for investigating the cellular roles of CARD19 without clonal selection, representing a bulk population of gene-edited cells suitable for a broad range of functional assays. As a target-gene disruption approach, it enables the study of CARD19’s impact on signaling pathways in a near-haploid genetic background, facilitating robust and reproducible experimental outcomes.
The host HAP1 cell line is a male, adherent, near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) line. Its near-haploid karyotype simplifies genetic manipulation and data interpretation, making it a preferred platform for functional genomics and CRISPR-based screens. HAP1 cells retain key signaling pathways, including NF-kappaB, and provide a physiologically relevant context for studying immune signaling and apoptosis in a leukemia-derived model. Their robust growth characteristics and suitability for high-throughput applications further enhance their utility in target validation and mechanistic studies.
CARD19 encodes a mitochondrion- and endoplasmic reticulum-localized protein that binds Bcl10 via its CARD domain, inhibiting Bcl10-mediated NF-kappaB activation and apoptosis. It sequesters Bcl10 from the CARD11?CBcl10?CMALT1 complex to prevent aberrant signaling. Upon stimulation by upstream regulators such as tumor necrosis factor (TNF), interleukin-1 (IL-1), or lipopolysaccharide (LPS), the inhibitory interaction is relieved, allowing Bcl10 to associate with the IKK complex (IKK-alpha, IKK-beta, IKK-gamma), leading to phosphorylation and degradation of IkappaB-alpha and subsequent nuclear translocation of NF-kappaB (p65/p50) dimers. This triggers transcription of pro-inflammatory cytokines including IL-6 and TNF-alpha. CARD19 thus functions as a checkpoint in innate immune signaling, and its disruption dysregulates cell survival and apoptotic programs.
In the HAP1 CML background, CARD19 knockout offers a model to dissect the interplay between oncogenic signaling and inflammatory responses. The near-haploid genome ensures that a single genetic alteration yields a clear phenotype, minimizing masking by wild-type alleles. This model is valuable for examining how loss of CARD19 influences Bcl10-dependent NF-kappaB activation in leukemia, potentially revealing vulnerabilities in CML survival. Moreover, the absence of CARD19-mediated inhibition may sensitize cells to apoptotic stimuli, facilitating drug sensitivity and resistance studies in inflammatory and autoimmune diseases.
Typical research applications include functional characterization of CARD19 using NF-kappaB luciferase reporter assays to quantify transcriptional activity, apoptosis assays via annexin V/PI flow cytometry, western blotting for phospho-p65 and cleaved caspases, co-immunoprecipitation of CARD19?CBcl10 complexes, and RT-qPCR for NF-kappaB target gene expression. These polyclonal knockout cells are also suited for drug target validation in inflammatory diseases and cancer, enabling the assessment of how CARD19 deficiency modulates responses to pathway inhibitors. For further details, including batch-specific editing efficiency and culture conditions, please contact Ascent Research.
