The CBFA2T3 Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population derived from the HAP1 cell line, engineered to disrupt the CBFA2T3 gene locus. This product provides a loss-of-function model for investigating the transcriptional corepressor CBFA2T3 in a near-haploid human cellular context. The polyclonal format yields a heterogeneous knockout population, circumventing the bottleneck of single-cell cloning while maintaining applicability for pooled functional genomics studies. This gene-edited pool is suitable for biochemical, genetic, and pharmacological assays that require robust depletion of CBFA2T3 protein expression without clonal selection.
HAP1 is a chronic myeloid leukemia (CML)-derived cell line originating from a male patient, exhibiting an adherent fibroblast-like morphology and a near-haploid karyotype for most chromosomes. This haploid configuration simplifies gene editing and genetic screening, as single-allele mutations directly manifest phenotypically. HAP1 cells retain key signaling pathways relevant to hematopoiesis and leukemogenesis, including functional TP53 and intact Rb pathway, while supporting efficient transfection and stable knockout generation. The near-haploid background ensures uniform gene dosage effects, making HAP1 a widely adopted platform for CRISPR-based screens and mechanistic dissection of cancer-relevant genes.
CBFA2T3 (MTG16/ETO2) is a transcriptional corepressor that operates within the NCOR1/SIN3A/HDAC complex to silence gene expression via histone deacetylation. It directly interacts with HDAC1, HDAC2, and NCOR1, and associates with hematopoietic transcription factors TAL1, LMO2, GATA1, and RUNX1. CBFA2T3 homodimerizes to scaffold corepressor assembly and is recruited to chromatin through these DNA-binding partners. It functions downstream of Notch and TGF??? signaling, and upstream of target genes CDKN1A and MYC, thereby linking external cues to cell cycle arrest and differentiation. Disruption of CBFA2T3 dismantles this repressive hub, lifting constraints on proliferation-associated transcripts.
In HAP1 cells, CBFA2T3 knockout abrogates corepressor function, derepressing CDKN1A and MYC and potentially altering proliferation and erythroid differentiation programs. The near-haploid background amplifies these effects by eliminating compensation from a second allele, facilitating clear phenotypic readouts. This model is therefore well-suited for studying the role of CBFA2T3 in leukemia biology, as its loss or dysregulation is observed in acute myeloid leukemia and myelodysplastic syndromes. The polyclonal population enables robust, reproducible experiments without clonal artifacts, and supports large-scale CRISPR modifier screens to uncover synthetic lethal partners or drug response modulators.
This product supports diverse experimental workflows including quantitative western blotting and RT?qPCR for CBFA2T3 and target genes, co-immunoprecipitation for corepressor complex integrity, ChIP?qPCR for promoter binding dynamics, and flow cytometric analysis of hematopoietic markers. Proliferation assays, transcriptome profiling by RNA?seq, and chemical perturbation studies further expand its utility. The CBFA2T3 Knockout HAP1 Polyclonal Cells are thus a versatile resource for dissecting transcriptional repression mechanisms in hematopoiesis, evaluating the dependence of leukemia cells on CBFA2T3, and advancing drug discovery for corepressor-associated malignancies. For further inquiries, please contact Ascent Research.
