The CD163 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the CD163 gene in the near-haploid human HAP1 cell line. This product provides a heterogeneous pool of edited cells, each carrying targeted gene disruptions within the CD163 locus, enabling loss-of-function studies without clonal selection. The polyclonal format preserves population-level diversity and is well-suited for pooled phenotypic screens, bulk biochemical assays, and comparative analyses where gene knockout effects are assessed across a mixed genetic background. Researchers can utilize these cells to interrogate CD163-dependent processes in a genetically tractable human myeloid model system.
HAP1 cells are derived from the KBM-7 chronic myeloid leukemia line and possess a near-haploid karyotype, which simplifies genetic manipulation and facilitates unambiguous genotype-phenotype correlations. As a hematopoietic cell line in blast crisis, HAP1 retains myeloid features and is widely employed in high-throughput genetic screens, drug-sensitivity profiling, and functional genomics. Their rapid growth and stable maintenance under standard culture conditions make them a practical host for CRISPR-based knockout experiments, particularly for genes involved in hematopoietic and immune functions.
CD163 encodes a type I transmembrane scavenger receptor predominantly expressed on macrophages. It binds hemoglobin-haptoglobin complexes with high affinity, mediating their endocytosis and subsequent lysosomal degradation. This process liberates heme, which is catabolized by heme oxygenase-1 (HO-1), triggering a cascade that includes interleukin-10 (IL-10) secretion, STAT3 activation, and PI3K/Akt signaling. The receptor also interacts with RAGE and TREM2, connecting it to broader immune regulatory networks. Upstream, CD163 expression is induced by IL-10, glucocorticoids, and IL-6, and its engagement promotes anti-inflammatory macrophage polarization and tissue remodeling.
In the HAP1 myeloid context, CD163 knockout serves as a valuable tool for dissecting hemoglobin clearance and iron metabolism pathways, as well as the cellular response to hemolytic stress. The model enables examination of how loss of CD163 alters downstream signaling through HO-1, IL-10, and STAT3, and its impact on macrophage polarization states. Given the relevance of CD163 to atherosclerosis, hemolytic disorders, sepsis, and tumor-associated macrophages, this knockout system provides a reductionist platform for mechanistic studies that can complement primary cell and in vivo models.
Typical applications include western blotting and flow cytometry to confirm CD163 protein ablation, RT-qPCR to assess transcriptional changes in downstream targets, and hemoglobin uptake assays to quantify scavenger function. Co-immunoprecipitation experiments can probe receptor-ligand interactions, while cytokine profiling (e.g., IL-10 ELISA) and RNA-seq enable comprehensive signaling and transcriptomic analyses. The cells are also amenable to phagocytosis assays and high-content imaging, supporting drug discovery efforts aimed at modulating macrophage activity in inflammatory diseases and cancer. For further details, please contact Ascent Research.
