The IFI27 Knockout HAP1 Polyclonal Cells product comprises a heterogeneous population of CRISPR/Cas9-edited HAP1 cells with targeted disruption of the IFI27 gene. This polyclonal knockout cell pool provides a ready-to-use loss-of-function model for investigating IFI27-mediated biological processes without the isolation of single-cell clones. The CRISPR/Cas9 genome editing approach introduces gene-disruptive modifications across the cell population, enabling functional studies of IFI27 in a near-haploid human cell background.
The HAP1 cell line is a near-haploid human chronic myeloid leukemia (CML) cell line derived from the KBM-7 line, originating from a patient in blast crisis. HAP1 retains a single copy of most chromosomes, except for a disomic region on chromosome 15, which simplifies genetic analysis and facilitates unambiguous genotype?Cphenotype correlations. As an immortalized myeloid cell line, HAP1 retains signaling pathways relevant to hematopoietic malignancies, including type I interferon and JAK-STAT cascades, making it a suitable host for studying interferon-inducible genes such as IFI27.
IFI27 encodes an interferon-inducible protein localized to mitochondria and the nuclear envelope, functioning at the intersection of antiviral innate immunity, apoptosis, and cell cycle regulation. Expression is driven by type I interferons (IFN-??/??) via the JAK-STAT axis, utilizing receptor IFNAR1, kinases JAK1 and TYK2, transcription factors STAT1 and STAT2, and cofactor IRF9. IFI27 promotes mitochondrial outer membrane permeabilization by interacting with Bcl-2 family members and mitochondrial proteins, releasing cytochrome c to activate caspase-3. Through p53-dependent pathways, IFI27 induces cell cycle inhibitors like p21, contributing to growth arrest. Thus, IFI27 translates interferon signals into pro-apoptotic and anti-proliferative outcomes.
In the HAP1 myeloid leukemia background, IFI27 knockout allows researchers to dissect its role in apoptosis sensitivity and interferon-mediated growth suppression. Given the near-haploid genome, the polyclonal knockout population provides a genetically homogenous system for screening modulators of IFI27-dependent pathways without clonal artifacts. This model is particularly valuable for examining how IFI27 loss influences chronic myeloid leukemia cell survival, drug responsiveness, and immune evasion mechanisms, potentially informing therapeutic strategies for hematological malignancies and solid tumors where IFI27 expression is dysregulated.
The IFI27 Knockout HAP1 Polyclonal Cells are suited for diverse experimental workflows, including interferon stimulation assays coupled with RT-qPCR or RNA-seq to profile downstream transcriptional changes, flow cytometric apoptosis analysis using Annexin V/PI staining, assessment of mitochondrial membrane potential with JC-1 dye, and caspase activity measurements. Immunofluorescence can confirm IFI27 subcellular localization, while co-immunoprecipitation enables mapping of protein interactions with Bcl-2 family members or nuclear pore components. Cell cycle analysis via propidium iodide staining and viability assays further support functional characterization. For further details and technical support, please contact Ascent Research.
