The EFR3B Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population specifically engineered to disrupt the EFR3B gene in the HAP1 human near-haploid cell line. This product provides a powerful loss-of-function model for investigating the role of EFR3B as a scaffold protein in phosphoinositide metabolism and PI4K complex signaling.
HAP1 is a human near-haploid cell line derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid genotype eliminates functional redundancy from a second allele, enabling clean genetic backgrounds for knockout studies. HAP1 is widely utilized in functional genomics and high-throughput screening due to its ease of genetic manipulation and robust growth characteristics.
EFR3B encodes a palmitoylated membrane anchor essential for the assembly of the phosphatidylinositol 4-kinase (PI4K) complex at the plasma membrane. Mechanistically, EFR3B recruits PI4KA via the TTC7 adaptor proteins (TTC7A and TTC7B) and interacts with FAM126A, facilitating the local synthesis of phosphatidylinositol 4-phosphate (PI4P). This lipid second messenger engages downstream effectors such as oxysterol-binding protein (OSBP) and FAPP1, and modulates AKT signaling. Upstream regulators include cell adhesion signals and growth factor receptors, positioning EFR3B at the interface between membrane microdomain organization and downstream signaling cascades that control membrane trafficking and cell adhesion.
In the HAP1 background, disruption of EFR3B perturbs PI4K complex formation and reduces plasma membrane PI4P levels, offering a tractable system to study its contribution to phosphoinositide-dependent pathways. Given the potential involvement in PI3K/AKT signaling and its interplay with cell adhesion, this knockout model is particularly relevant for cancer biology and signal transduction research. The near-haploid nature further enables synthetic lethal screens and modifier studies to identify functionally related genes and drug targets.
Researchers can employ this model in a variety of assays, including Western blotting for PI4P quantification, immunofluorescence to visualize plasma membrane lipid distribution, phospho-AKT ELISA to measure signaling output, cell migration assays, and flow cytometry for integrin expression profiling. The polyclonal format is compatible with pooled high-throughput genetic screens and metabolic activity analyses, making it a versatile tool for both mechanistic studies and drug discovery campaigns. For further details, please contact Ascent Research.
