The GPSM2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the GPSM2 gene has undergone targeted disruption. Unlike monoclonal lines, this polyclonal pool encompasses a spectrum of edited alleles, providing a robust loss-of-function model for bulk functional assays and pooled genetic screens where clonal homogeneity is not required.
The HAP1 host cell line is a near-haploid, fibroblast-like human cell line derived from the KBM-7 chronic myeloid leukemia line, exhibiting adherent morphology. Its near-haploid genetic context facilitates efficient CRISPR/Cas9-mediated gene disruption and reduces the complication of a functional wild-type allele, thereby enhancing the clarity of phenotypic changes observed in knockout populations.
GPSM2 (also known as LGN) is a pivotal regulator of G protein signaling that directs mitotic spindle orientation and asymmetric cell division. It is recruited to the cell cortex by interacting with G??i??GDP (GNAI1/GNAI2/GNAI3), where it scaffolds NuMA and cytoplasmic dynein to generate pulling forces on astral microtubules. This cortical G??i??GPSM2-NuMA-dynein complex is modulated by upstream polarity factors, including Frizzled/Dishevelled and the adaptor INSC, linking it to Wnt/planar cell polarity pathways. Furthermore, GPSM2 interacts with DLG1 and the microtubule depolymerase KIF2A and has been shown to regulate the Hippo signaling cascade through LATS1/2 kinases, thereby influencing cortical actin dynamics and downstream gene transcription.
In the HAP1 near-haploid background, disruption of GPSM2 provides an uncomplicated model for dissecting the molecular underpinnings of asymmetric division and spindle orientation. The lack of a compensatory diploid allele allows for unambiguous assessment of GPSM2??s role in processes such as G protein-coupled receptor-mediated spindle control and planar cell polarity-dependent division. This model is especially pertinent to research on autosomal recessive nonsyndromic hearing loss DFNB82 and Chudley-McCullough syndrome, where GPSM2 mutations may disrupt asymmetric cell fate decisions. It also serves as a valuable tool in cancer biology for exploring the consequences of defective spindle orientation on tumor cell division.
Researchers can employ this polyclonal knockout model in diverse experimental settings. It is well suited for immunofluorescence-based analysis of spindle orientation by visualizing NuMA and dynein distribution, and for live-cell imaging of mitotic progression. The cells can be used in pooled genetic screens to discover novel regulators of asymmetric division, and in biochemical assays such as co-immunoprecipitation to verify GPSM2-containing complexes. Standard techniques including western blotting for protein-level knockout confirmation, qPCR for assessing target gene expression changes, and flow cytometry for cell cycle profiling are directly applicable. For additional technical specifications and ordering details, please contact Ascent Research.
