The CD14 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population with ablated CD14 expression, derived from the near-haploid HAP1 human cell line. This polyclonal knockout model provides a genetically diverse pool of cells for robust loss-of-function studies of CD14, a glycosylphosphatidylinositol (GPI)-anchored co-receptor for bacterial lipopolysaccharide (LPS) critically involved in innate immune recognition.
The HAP1 cell line is a near-haploid human line derived from KBM-7 chronic myeloid leukemia (CML) cells. Its haploid genome simplifies gene editing and genotype-phenotype correlations, making it a powerful tool for functional genomics and CRISPR-based genetic screens. HAP1 cells retain myeloid lineage characteristics, providing a physiologically relevant context for investigating innate immune signaling pathways involving CD14.
CD14 is a high-affinity LPS receptor that facilitates LPS transfer to TLR4/MD-2, triggering MyD88- and TRIF-dependent signaling. This leads to activation of IRAK1/4, TRAF6, and TAK1, which phosphorylate the IKK complex and MAPKs, resulting in NF-??B, AP-1, and IRF3 activation and production of TNF-??, IL-6, IL-1??, and type I interferons. CD14 expression is induced by LPS, IFN-??, and TNF-?? via Sp1, Sp3, and C/EBP transcription factors. CD14 functions in concert with LBP and TIRAP/Mal at the TLR4 complex and mediates phagocytosis of bacterial pathogens.
In the HAP1 host background, CD14 knockout provides a clean loss-of-function model to dissect TLR4-mediated innate immune responses. The near-haploid genome allows efficient knockout of additional pathway components for epistasis studies and simplifies genetic interaction screens. Because HAP1 retains myeloid characteristics, this model is physiologically apt for studying LPS-induced inflammation and the role of CD14 in pathogen recognition. The polyclonal nature reduces clonal artifacts, yielding robust population-level data.
The CD14 Knockout HAP1 Polyclonal Cells are suited for a variety of functional assays, including flow cytometry to confirm CD14 surface loss, NF-??B reporter assays for pathway activity, Western blotting for phospho-NF-??B and phospho-p38, and ELISA for TNF-?? and IL-6 quantification. These cells enable research on innate immunity, LPS/TLR4 signaling, anti-inflammatory drug screening, host-pathogen interactions, and genome-wide CRISPR screens. RNA-seq can be used to profile transcriptomic alterations due to CD14 deficiency. For further technical details, please contact Ascent Research.
