The GSPT2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the GSPT2 locus has been disrupted to abolish its function. This pooled polyclonal format provides a robust loss-of-function model for investigating the biological role of the translation termination factor eRF3 in a near-haploid background. The product is supplied as a heterogeneous population of edited cells, allowing researchers to bypass clonal selection while maintaining genetic knockout across the culture.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its hemizygous genome simplifies CRISPR/Cas9-mediated gene targeting and enables the study of recessive phenotypes without the confounding effects of a second allele. Widely used in functional genomics and drug discovery, HAP1 cells retain key signaling pathways relevant to leukemia and cancer research while offering an experimentally tractable system for genetic manipulation.
GSPT2 encodes eRF3a, a GTPase that cooperates with eRF1 to mediate translation termination at stop codons. Upon GTP hydrolysis, eRF3a promotes peptide release and ribosomal subunit dissociation, thereby regulating global protein synthesis. Beyond its canonical role in translation, GSPT2 contributes to G1/S cell cycle progression, likely through modulating the translation of cell cycle regulators. Upstream, GSPT2 expression is controlled by E2F transcription factors and cyclin-dependent kinases, linking it to proliferation signals. It interacts directly with eRF1 and ribosomal subunits, and its function is integrated within the translation termination machinery alongside eRF1 and the ribosome.
In the haploid HAP1 background, knockout of GSPT2 eliminates eRF3a activity without interference from a functional second allele, enabling clean dissection of its roles in translation termination and cell cycle control. This model is particularly suited for studying the consequences of impaired termination fidelity, such as readthrough or premature termination, and for assessing how altered protein synthesis affects the G1/S transition. The near-haploid state also facilitates genetic screens for synthetic lethal interactions or modulators of GSPT2-dependent pathways in leukemia contexts.
Typical applications include western blotting to confirm loss of GSPT2 protein, RT-qPCR for mRNA analysis, flow cytometry for cell cycle profiling, polysome profiling to evaluate translation activity, and luciferase reporter assays to measure termination fidelity. These cells also enable functional genomics studies in haploid cells, cancer cell cycle research, and investigation of translational control mechanisms. For further technical information or custom cell line services, please contact Ascent Research.
