The EID3 Knockout HAP1 Polyclonal Cells product consists of a polyclonal population of HAP1 cells engineered with CRISPR/Cas9-mediated disruption of the EID3 locus, generating a loss-of-function model to interrogate EID3-dependent transcriptional and cell cycle control. This polyclonal knockout pool enables the study of heterogeneous gene-disruption outcomes without the clonal bias inherent to single-cell-derived lines, making it well-suited for pooled functional screens and population-level analyses. The CRISPR/Cas9 approach introduces targeted genomic lesions that abolish EID3 protein expression, thereby recapitulating corepressor deficiency in a human cell context.
The parental HAP1 cell line is a near-haploid, adherent cell line originally derived from the KBM-7 chronic myelogenous leukemia (CML) cell line. Its near-haploid karyotype simplifies genetic manipulation and facilitates high-quality CRISPR knockout experiments by reducing gene-copy redundancy and minimizing compensatory effects from homologous alleles. HAP1 cells maintain a stable, near-haploid state for many core chromosomes, making them an established platform for functional genomics screens, drug sensitivity profiling, and mechanistic studies in hematological malignancy models.
EID3 functions as a transcriptional corepressor through direct physical interaction with the histone acetyltransferases EP300 (p300) and CREBBP (CBP), inhibiting their acetyltransferase activity. This interaction reduces histone acetylation at regulatory regions of target genes, thereby repressing transcription. EID3 is activated downstream of the Notch intracellular domain (NICD) and the E2A-PBX1 fusion oncogene, and it dampens expression of Notch target genes such as HES1 and HEY1. It also modulates cell cycle progression by repressing genes including Cyclin D1, positioning EID3 at the intersection of differentiation signals and proliferative control.
In the near-haploid HAP1 background, loss of EID3 eliminates a key node in p300/CBP-dependent transcriptional regulation, allowing researchers to dissect the corepressor??s contribution to leukemogenic programs. The knockout model in a CML-derived lineage provides a reductionist system for studying EID3??s role in myeloid malignancies, where E2A-PBX1 and Notch signaling are often dysregulated. The absence of a second allele in many loci magnifies the phenotypic impact of EID3 disruption, enhancing the sensitivity of downstream assays and reducing confounding genetic complementation.
Typical experimental applications include assessing global transcriptional changes via RNA-seq, validating EID3 protein loss by Western blot, and performing co-immunoprecipitation to confirm disrupted EP300/EID3 complexes. The polyclonal population is amenable to drug sensitivity profiling using viability assays such as MTT, enabling the identification of EID3-dependent therapeutic vulnerabilities. These cells also support pooled CRISPR modifier screens and cell cycle analyses. For detailed product specifications or ordering assistance, please contact Ascent Research.
