The EAF2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that provides targeted disruption of the EAF2 gene in the HAP1 human near-haploid cell line. This loss-of-function model abolishes the expression of EAF2, a critical subunit of the super elongation complex (SEC), which is essential for productive RNA polymerase II (Pol II)-dependent transcription elongation. The polyclonal nature of this knockout product preserves genetic diversity while uniformly eliminating EAF2 protein function, making it ideal for population-based functional assays where clonal variation is not desired.
The HAP1 host cell line is a near-haploid, adherent, fibroblast-like cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype simplifies genetic analysis and knockout generation, as only one allele requires targeting to achieve functional gene disruption. HAP1 cells retain intact p53 signaling and key transcription pathways, making them a robust system for investigating tumor suppressor functions and transcriptional regulation. The cells are widely used in genetic screens, drug sensitivity profiling, and mechanistic studies due to their stable growth and compatibility with various molecular assays.
EAF2 (ELL-associated factor 2) acts as a scaffold within the SEC, interacting directly with elongation factors ELL and AFF4, as well as the positive transcription elongation factor b (P-TEFb) kinase subunit CDK9 and its regulatory partner Cyclin T1. EAF2 transcription is activated by the tumor suppressor p53 and can be modulated by androgen receptor signaling. Downstream, functional SEC containing EAF2 promotes the expression of key immediate early genes such as MYC and FOS, the cell cycle inhibitor CDKN1A (p21), and the pro-apoptotic factor BAX. Disruption of EAF2 therefore impairs SEC assembly and processivity, leading to reduced transcriptional output at these loci. Consequently, knockout cells exhibit defective induction of elongation-dependent genes, which has been implicated in altered cell cycle control and apoptosis resistance.
In the HAP1 cell context, loss of EAF2 creates a powerful model to dissect SEC-dependent transcriptional programs without the confounding effects of genetic redundancy found in diploid lines. The near-haploid background ensures a clear link between EAF2 ablation and phenotype, facilitating gene dosage studies and synthetic lethality screens. This system is particularly relevant for studying prostate cancer and leukemia biology, where EAF2 functions as a tumor suppressor. The knockout model can be employed to investigate how EAF2 integrates signals from upstream regulators like p53 and androgen receptor to control proliferation and survival pathways.
Typical applications include transcriptional regulation studies via RNA-seq and RT-qPCR, assessment of apoptosis and cell cycle by flow cytometry, protein interaction analyses by co-immunoprecipitation, and functional screening in drug sensitivity assays. Researchers can monitor downstream target expression (e.g., MYC, FOS, CDKN1A) by Western blotting to validate pathway engagement. The cells are also suited for high-throughput screens to identify synthetic lethal partners or compounds that selectively affect EAF2-deficient cancers. For additional information or custom inquiries, please contact Ascent Research.
