The IFRD1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population with disruption of the IFRD1 gene in the HAP1 near-haploid human cell line. This loss-of-function model enables investigation of IFRD1-dependent transcriptional regulation and cellular differentiation. The polyclonal format provides a heterogeneous mixture of edited alleles, suitable for bulk population studies. It serves as a tool for functional genomics, drug target validation, and hematopoietic biology research.
HAP1 is a near-haploid cell line derived from KBM-7 chronic myeloid leukemia cells. Its haploidy facilitates gene-editing and homozygous knockout generation, making it ideal for genetic screens and genotype-phenotype studies. As a hematopoietic model, it retains myeloid lineage features, and the reduced genetic redundancy simplifies loss-of-function analyses, aiding dissection of signaling pathways in leukemogenesis and differentiation.
IFRD1 functions as a transcriptional corepressor by directly binding histone deacetylases HDAC1 and HDAC2, along with TLE corepressor proteins, to mediate histone deacetylation and long-range gene silencing. It represses key target genes, including the myogenic factor MyoD and genes essential for myeloid differentiation. Upstream, IFRD1 is activated by PKC in response to TPA and acts as a downstream effector of Notch signaling (via NICD, RBP-J, and HES1) and TGF-beta pathways. It is under direct transcriptional control by the hematopoietic master regulator RUNX1 and cooperates with MEF2C to coordinate differentiation, thereby integrating multiple signaling inputs.
In HAP1, IFRD1 knockout disrupts corepressor complexes, potentially derepressing proliferation and differentiation genes. As a RUNX1 target gene implicated in AML and MDS, this model is valuable for studying leukemogenesis mechanisms. The haploid background minimizes paralog interference, and the polyclonal pool captures diverse mutations, useful for pooled screens or drug sensitivity assays assessing epigenetic therapies.
Researchers can perform Western blotting and RT-qPCR to confirm knockout and target derepression, RNA-seq for transcriptome profiling, and flow cytometry for myeloid differentiation markers. Co-immunoprecipitation and ChIP-qPCR assess repressive complex integrity and histone acetylation. Luciferase reporters monitor pathway activity, and HDAC inhibitor sensitivity assays explore therapeutic vulnerabilities. This reagent advances transcriptional regulation studies in hematopoiesis. Contact Ascent Research for details.
