The CD1A Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 human cell line, designed for loss-of-function studies of CD1A. Gene disruption results in ablation of CD1A protein expression, and the polyclonal format provides a heterogeneous pool of edited cells, offering a reliable model for functional assays.
HAP1 is a near-haploid human cell line originating from a male patient with chronic myeloid leukemia (CML); it displays fibroblast-like morphology and is a cornerstone for genetic screening and functional genomics due to its haploid karyotype. This characteristic ensures that knockout of a single allele leads to a null phenotype, facilitating robust and interpretable experiments. The line??s ease of transfection and stable growth further support CRISPR-mediated gene editing and downstream analyses.
CD1A encodes a lipid antigen-presenting molecule that forms heterodimers with beta-2 microglobulin (B2M) and relies on saposin C for loading lipid antigens. Expression is regulated by cytokines GM-CSF, IL-4, TGF-??, and the transcription factor PU.1. Upon recognition by the TCR, the CD1A?Clipid complex triggers the Src-family kinase Lck, which phosphorylates ZAP70, leading to phosphorylation of the adaptor LAT and activation of NF-??B, NFAT, and AP-1. These transcription factors drive the expression of cytokines such as IFN-??, IL-4, and IL-17, promoting T cell differentiation and effector functions.
In the near-haploid HAP1 background, CRISPR-mediated CD1A disruption abolishes lipid antigen presentation to CD1a-restricted T cells, impairing T cell priming, cytokine secretion, and antimicrobial activity. This clean genetic system enables dissection of CD1A-dependent pathways in antigen presentation and inflammation.
Researchers can validate CD1A knockout by flow cytometry or immunofluorescence, then perform co-culture assays with CD1a-restricted T cell clones to measure cytokine output via ELISA. Lipid antigen loading experiments coupled with T cell proliferation assays clarify presentation requirements, while bacterial killing assays provide functional readouts of antimicrobial responses. RNA-seq can identify transcriptomic changes resulting from CD1A loss. These approaches facilitate studies in host-pathogen interactions, autoimmune diseases, and drug discovery for immune modulators. For further details, contact Ascent Research.
