GRB10 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 haploid human cell line. This product provides a heterogeneous pool of cells with targeted disruption of the GRB10 gene, enabling loss-of-function studies in a near-haploid genetic background.
The HAP1 cell line is a haploid fibroblast-like leukemia cell line originally derived from a male patient with chronic myeloid leukemia. Its near-haploid karyotype simplifies genetic manipulation and phenotypic analysis, as only one allele must be targeted to achieve functional knockout. This makes HAP1 an ideal host for CRISPR/Cas9-mediated gene disruption, particularly for studying gene function in signal transduction and metabolic regulation.
GRB10 encodes an adaptor protein that negatively regulates insulin and insulin-like growth factor 1 (IGF-1) receptor signaling. It binds to activated insulin and IGF-1 receptors via its SH2 domain, competitively inhibiting the recruitment of insulin receptor substrate 1 (IRS1) and attenuating downstream PI3K-Akt and mTOR signaling. GRB10 further suppresses pathway output by recruiting the NEDD4 ubiquitin ligase to promote receptor ubiquitination and degradation. This polyclonal knockout model disrupts GRB10 expression, thereby relieving its inhibitory effects on insulin and IGF-1 receptor cascades and enhancing signaling through Akt, mTOR, and S6K.
In the HAP1 background, GRB10 knockout cells offer a simplified system to dissect insulin and growth factor signaling without the complexity of a diploid genome. The haploid state ensures that gene disruption in the polyclonal population effectively eliminates functional GRB10 protein, facilitating consistent phenotypic outcomes. Researchers can explore how loss of this adaptor alters receptor stability, signal duration, and downstream metabolic responses.
These cells are suitable for investigating insulin resistance mechanisms, cancer-related growth factor signaling, and metabolic disease modeling. Typical applications include western blot analysis of insulin receptor phosphorylation and total levels, co-immunoprecipitation to study GRB10-receptor interactions, glucose uptake assays, and phospho-Akt flow cytometry. By providing a straightforward knockout model in a haploid cell platform, GRB10 Knockout HAP1 Polyclonal Cells accelerate functional studies linking insulin and IGF-1 receptor pathways to cellular metabolism and proliferation. For further information, please contact Ascent Research.
