The BATF3 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting BATF3 in HAP1 cells. This heterogeneous pool harbors diverse loss-of-function mutations, avoiding clonal bias and enabling robust population-level functional screens in a near-haploid background. The model retains HAP1 characteristics while ablating BATF3, suitable for dissecting dendritic cell biology and immune regulation.
HAP1 is a near-haploid adherent cell line derived from the KBM-7 chronic myeloid leukemia line. Its reduced genomic complexity facilitates CRISPR-mediated gene disruption and minimizes false-positive effects from recessive mutations, making it a premier model for haploid genetic screens and functional genomics. Its hematopoietic origin renders it particularly relevant for studying immune cell development and signaling pathways.
BATF3 encodes a lineage-determining basic leucine zipper transcription factor essential for CD8??+ conventional dendritic cell (cDC1) development and cross-presentation to CD8+ T cells. It cooperates with IRF8 and AP-1 components JUN and BATF, downstream of Flt3 ligand and GM-CSF receptor activation, regulated by STAT3/STAT5 and PU.1. BATF3 transactivates IL12B, XCR1, CLEC9A, and TLR3, and engages in feedforward regulation with IRF8, integrating Toll-like receptor signals to drive IL-12 production and adaptive immunity.
In the HAP1 background, BATF3 disruption offers a simplified system to dissect the molecular requirements for cDC1 lineage commitment. The hematopoietic origin of the cells provides a relevant context for probing the Flt3?CIRF8?CBATF3 axis, and the knockout phenotype enables interrogation of antigen cross-presentation defects, altered cytokine profiles, and impaired T cell activation. This reductionist model facilitates mapping of BATF3-dependent transcriptional targets without the confounding variables of primary dendritic cell cultures.
Researchers can employ this polyclonal knockout pool in western blotting to confirm BATF3 loss, RT-qPCR to quantify downstream effectors like IL12B, and flow cytometry to assess cDC1-associated markers XCR1 and CLEC9A. Functional co-culture assays with CD8+ T cells measure cross-presentation efficiency, and multiplex cytokine assays detect IL-12 secretion. RNA-seq reveals transcriptomic changes. Applications span tumor immunology, vaccine adjuvant research, antiviral immunity, and CRISPR-based functional genomics. For further details, please contact Ascent Research.
