The GOPC Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population in which the GOPC gene has been disrupted to create a loss-of-function model for studying Golgi-associated scaffold protein function. This polyclonal knockout product contains a heterogeneous pool of edited cells, providing a robust system for investigating the cellular consequences of GOPC deficiency without the clonal variability associated with single-cell-derived lines. The use of CRISPR/Cas9-mediated gene disruption ensures efficient targeting of the GOPC locus, enabling researchers to dissect its role in intracellular trafficking and signal transduction.
The host cell line, HAP1, is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia (CML) line. Its haploid karyotype simplifies genetic manipulation and phenotypic analysis, as a single gene disruption can produce a complete loss-of-function phenotype without the confounding effects of a second allele. HAP1 cells are widely employed in genetic screens, knockout validation, and receptor signaling studies, making them an ideal background for investigating GOPC-dependent processes. The CML origin also provides a cancer-relevant context, particularly for pathways dysregulated in leukemia and other malignancies.
GOPC encodes a PDZ domain-containing scaffold protein at the trans-Golgi network that is essential for post-Golgi trafficking of receptors like EGFR, TGFBR, and Frizzled. GOPC functions downstream of activated EGFR, facilitating receptor recycling to the plasma membrane to sustain signaling. It also interacts with CFTR, BAI1, Grb2, and CLCN3 to coordinate vesicular transport and actin reorganization. In Wnt signaling, GOPC regulates Frizzled trafficking, affecting ??-catenin stabilization via GSK3B and ACTB. Thus, GOPC disruption alters surface expression of key receptors, impairing EGFR, TGF-??, and Wnt pathway outputs.
In the HAP1 background, GOPC knockout leads to profound trafficking defects that can be readily characterized using the haploid genetic system. The absence of a second functional allele ensures that observed phenotypes stem directly from GOPC loss, enhancing the clarity of genotype?Cphenotype correlations. This model is particularly valuable for dissecting the scaffolding functions of GOPC in cancer-related signaling, as HAP1 cells retain key signaling modules such as EGFR, TGFBR, and Wnt components that are frequently deregulated in CML and solid tumors. Moreover, the CML derivation connects GOPC study to hematopoietic malignancies, where receptor recycling and Golgi dynamics contribute to oncogenic signaling.
Applications of GOPC knockout HAP1 polyclonal cells include dissecting receptor trafficking mechanisms using immunofluorescence microscopy for Golgi markers, western blotting for target protein levels, and receptor internalization assays. Flow cytometry can quantify changes in surface receptor expression, while drug sensitivity screens enable identification of compounds with synthetic lethal interactions in GOPC-deficient cancer cells. These cells provide a versatile platform for studying signal transduction in a haploid background, supporting cancer cell biology and drug discovery research. For additional information, please contact Ascent Research.
