The CD1E Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population of HAP1 cells, genetically disrupted at the CD1E locus. This product provides a heterogeneous pool of cells carrying diverse loss-of-function mutations, generated without clonal isolation, preserving polyclonal diversity for functional studies. Researchers obtain a ready-to-use knockout model that abolishes CD1E expression and function, enabling dissection of CD1E-dependent lipid antigen presentation in a human near-haploid background.
HAP1 is a near-haploid, fibroblast-like cell line derived from KBM-7 chronic myeloid leukemia cells. It features a predominantly haploid genome and p53 deficiency, which facilitates efficient CRISPR-based gene disruption and reduces DNA damage-induced cell cycle arrest. This makes HAP1 an ideal host for large-scale genetic screens, high-throughput functional genomics, and targeted knockout studies, as single-copy disruptions often yield full loss-of-function phenotypes without the need for biallelic targeting.
CD1E is an MHC class I-like molecule that presents lipid antigens to T cells. It forms complexes with ??2-microglobulin (B2M) and localizes to lysosomes, where cathepsin L and saposin C facilitate lipid loading. Upstream, CD1E expression is induced by GM-CSF, IL-4, and TLR ligands via transcription factor PU.1. Downstream, CD1E-mediated antigen presentation activates TCR signaling, promoting IFN-?? and IL-4 secretion, linking innate immune triggers to adaptive T cell responses.
In the HAP1 setting, CD1E knockout disrupts the entire lipid antigen presentation pathway, providing a powerful system to interrogate CD1-restricted immunity. The near-haploid background ensures unambiguous genotype-phenotype correlations, while p53-deficiency removes apoptotic confounds common in primary cells, enhancing assay robustness. This model is valuable for investigating mycobacterial lipid presentation in infectious diseases like tuberculosis, autoantigen presentation in autoimmunity, and lipid-based vaccine strategies.
These polyclonal knockout cells support diverse applications, including haploid genetic screens to identify novel lipid-processing machinery, T cell activation assays with CD1E-restricted clones, and mechanistic studies of lysosomal lipid trafficking. Routine validation uses Western blotting, RT-qPCR, and flow cytometry for CD1E loss, while co-immunoprecipitation of B2M, immunofluorescence for lysosomal markers, and TIDE assay confirm functional knockout. Transcriptomic analysis (RNA-seq) can reveal pathway-wide adaptations. For inquiries, contact Ascent Research.
