The EEF1A2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the EEF1A2 gene in HAP1 cells. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous mix of edited alleles without clonal selection. It provides a versatile system for investigating EEF1A2-dependent translation elongation and associated cellular processes.
HAP1 is a near-haploid fibroblast-like cell line derived from the KBM-7 chronic myeloid leukemia isolate. Its haploid genome, except for a disomic chromosome 15, simplifies genetic manipulation and enables high-efficiency CRISPR targeting, making it a workhorse for functional genomics and drug screening. HAP1 retains key signaling networks relevant to cancer and neurodevelopment, offering a physiologically informed platform for gene knockout studies.
EEF1A2 encodes a translation elongation factor that delivers aminoacyl?tRNA to the ribosomal A?site, a critical step in mRNA translation that governs global protein synthesis. Beyond translation, EEF1A2 acts as an actin?bundling protein and inhibits apoptosis, in part by stabilizing Bcl?xL. Its expression is tightly controlled by oncogenic signals: MYC drives transcription, while mTORC1 downstream of PI3K/AKT modulates its activity. Additional regulatory input comes from ERK and STAT3 pathways. EEF1A2 directly interacts with actin, the eEF1B complex, and the ribosome, enabling coordinated regulation of protein synthesis and the cytoskeleton.
In the HAP1 knockout cells, loss of EEF1A2 disrupts elongation and re-organizes the actin network, making this model valuable for dissecting EEF1A2-associated pathologies. Mutations in EEF1A2 are linked to severe neurodevelopmental disorders, including intellectual disability, early?onset epilepsy, and autism spectrum disorder, while its overexpression in multiple cancers promotes tumor growth and chemoresistance. The haploid genome of HAP1 ensures that CRISPR targeting usually achieves complete gene inactivation, yielding a clean loss-of-function phenotype suitable for mechanistic studies.
Researchers can utilize these polyclonal knockout cells for diverse assays, including puromycin incorporation and polysome profiling to monitor translation rates, apoptosis and viability assays to probe survival signaling, and immunofluorescence to visualize actin reorganization. The polyclonal population is amenable to pooled CRISPR screens, drug screening, and genetic suppressor/enhancer screens, exploiting haploid genetics for unbiased discovery. Routine validation with western blotting, RT?qPCR, and flow cytometry confirms target knockout and pathway modulation. For additional technical information, please contact Ascent Research.
