The IL27 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the IL27 gene in the HAP1 human cell line. This format yields a heterogeneous pool of independently edited cells, each harboring distinct genetic modifications, thereby capturing the full breadth of CRISPR-induced variation. Such polyclonal populations are particularly advantageous for pooled functional genomics studies and avoid artifacts associated with single-clone selection.
HAP1 is a fibroblast-like, near-haploid human cell line derived from a male patient with chronic myelogenous leukemia. Its near-haploid karyotype eliminates allele heterozygosity, markedly simplifying gene editing, phenotypic analysis, and genotype interpretation. HAP1 cells exhibit robust growth, adherent morphology, and are widely adopted for CRISPR screens, protein interaction studies, and signaling pathway dissection, making them an ideal chassis for knockout model generation.
IL-27, a heterodimeric cytokine of the IL-12 family, comprises p28 and EBI3 subunits. It activates a receptor complex composed of IL27RA and the shared gp130 chain, stimulating receptor-associated JAK1, JAK2, and TYK2 kinases. This triggers phosphorylation and nuclear translocation of STAT1 and STAT3, initiating transcriptional programs central to immune regulation. Downstream targets include T-bet (driving Th1 differentiation), the immunosuppressive cytokine IL-10, the checkpoint molecule PD-L1, and the negative feedback regulator SOCS1. Upstream, IL-27 production is induced by Toll-like receptor agonists, IFN-??, and CD40 ligation, placing IL-27 at the intersection of innate and adaptive immunity.
Disrupting IL27 in the HAP1 background offers a powerful reductionist system to interrogate IL-27?Cdependent mechanisms. The near-haploid state ensures that knockout genotypes translate directly to phenotypes without background allelic interference. The polyclonal composition facilitates unbiased functional genetic screens, such as identifying synthetic lethal interactions or resistance mechanisms within the JAK-STAT network. This model is especially relevant for studying how IL-27 loss alters the balance between pro-inflammatory (Th1) and anti-inflammatory (IL-10) pathways, with implications for autoimmune disorders and tumor immune evasion.
These polyclonal knockout cells support a wide spectrum of experimental workflows. Routine validation of gene disruption can be performed via western blotting and RT-qPCR, while functional studies benefit from phospho-STAT1/3 flow cytometry to assess signaling dynamics and cytokine ELISA to measure IL-10 secretion. Transcriptomic approaches such as RNA-seq enable genome-wide expression profiling, and co-culture assays can model IL-27-mediated intercellular communication. Additionally, the cells serve as a robust backbone for arrayed or pooled CRISPR screens focusing on immune signaling. For project-specific assistance, contact Ascent Research.
