The CBFA2T2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the CBFA2T2 gene has been disrupted to generate a loss-of-function model. This product consists of a genetically heterogeneous pool of HAP1 cells carrying diverse CRISPR-induced mutations at the CBFA2T2 locus, providing a robust system for studying CBFA2T2 function without clonal selection bias.
HAP1 is a near-haploid human cell line with an adherent fibroblast-like morphology, originally derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype simplifies gene disruption, as single guide RNAs can effectively target alleles in haploid regions, making it an established platform for functional genomics, haploid genetic screening, and rapid knockout generation. This background is particularly relevant for myeloid biology research, given its leukemia origin.
CBFA2T2 encodes a transcriptional corepressor that represses gene expression by recruiting histone deacetylases (HDAC1, HDAC2) and the SIN3A/NCOR1 complex to promoters. DNA-binding factors such as RUNX1, TCF4, and LEF1 mediate its recruitment to target genes, where it silences loci including RUNX1 targets (IL3, CSF2), Wnt pathway genes (MYC, CCND1), and CDKN1A. CBFA2T2 activity is regulated by upstream signals from NOTCH1 and TCF3/TCF4, and it interacts with hematopoietic factors like TAL1. Through these interactions, CBFA2T2 integrates Wnt, Notch, and TGF-?? signaling to control hematopoietic proliferation and differentiation.
The myeloid origin of HAP1 cells and the established role of CBFA2T2 as a negative regulator of hematopoiesis and a putative tumor suppressor in myeloid malignancies make this knockout model highly relevant for leukemogenesis research. Disruption of CBFA2T2 in this context permits investigation of aberrant transcriptional programs driving AML, MDS, and therapy-related myeloid neoplasms. The polyclonal format avoids clonal artifacts, enabling robust evaluation of phenotypes such as altered proliferation, apoptosis, and differentiation.
These polyclonal knockout cells support a wide array of experimental applications. Researchers can assess CBFA2T2-dependent gene expression changes via RNA-seq and RT-qPCR, or examine chromatin modifications at target promoters using ChIP-qPCR for histone acetylation. Protein interaction studies with co-immunoprecipitation can probe associations with RUNX1, TCF4, and HDAC complexes. Functional assays including cell proliferation, apoptosis, and flow cytometric analysis of differentiation markers provide insights into CBFA2T2’s role in hematopoiesis. Wnt/Notch reporter assays allow exploration of pathway crosstalk, and the population is amenable to high-throughput CRISPR screens for modifier genes or drug target validation. For further information, please contact Ascent Research.
