EIF5A2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population in which the EIF5A2 gene has been disrupted via genome editing. This product provides a heterogeneous knockout model derived from the HAP1 near-haploid human cell line, offering a loss-of-function system to interrogate the biological roles of the eukaryotic translation initiation factor 5A2. The polyclonal format captures a range of editing outcomes across the cell population, making it suitable for pooled functional genomics screens and studies requiring a bulk knockout context without clonal isolation.
The HAP1 host cell line is a near-haploid chronic myeloid leukemia (CML) model originally derived from the KBM-7 cell line, representing a male genetic background. HAP1 cells retain a largely haploid karyotype, which simplifies the generation of bi-allelic gene disruptions through CRISPR/Cas9 editing and reduces genetic redundancy. These cells exhibit both adherent and suspension growth capabilities, offering versatility for various culture formats and high-throughput screening platforms. The HAP1 line has been widely adopted as a robust system for functional genomics, particularly for CRISPR-based knockout screens, due to the ease of achieving complete gene inactivation in a single allele.
EIF5A2 encodes a translation elongation factor that undergoes hypusination, a unique post-translational modification dependent on spermidine and catalyzed by deoxyhypusine synthase (DHPS) and deoxyhypusine hydroxylase (DOHH). The hypusinated protein associates with ribosomes to facilitate translation of specific proline-rich motif-containing mRNAs, including cyclin D1 and BCL-2 family members, thereby promoting cell cycle progression and inhibiting apoptosis. EIF5A2 expression is directly activated by the MYC transcription factor and responsive to spermidine levels, integrating its function into the MYC transcriptional network and mTOR signaling pathway. Its deregulation is implicated in multiple human malignancies.
The near-haploid nature of HAP1 cells ensures that CRISPR-mediated disruption of the single EIF5A2 allele results in a complete loss of functional protein, eliminating potential compensatory effects from a second allele. This genetic simplicity makes the knockout model particularly powerful for delineating EIF5A2-dependent processes in a cancer cell context, as the parental HAP1 line originates from a CML patient. The knockout population serves as a valuable tool to investigate how ablation of the hypusination-dependent translation elongation function affects global protein synthesis, cell proliferation, and apoptotic regulation, especially under conditions that mimic oncogenic stress or therapeutic interventions targeting the hypusine pathway.
This polyclonal EIF5A2 knockout model is applicable to a wide range of cancer biology and translation research studies. Standard characterization includes Western blotting for total and hypusinated EIF5A2, RT-qPCR, and functional assays for proliferation, apoptosis, cell cycle, and migration. Advanced applications encompass translatome analysis by RNA-seq, MYC reporter assays, co-immunoprecipitation of EIF5A2 interactors, and drug sensitivity testing with hypusination inhibitors like GC7 or deferiprone. For further information or to place an order, please contact Ascent Research.
