The ARFGAP2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ARFGAP2 gene in human HAP1 cells. This loss-of-function model is generated by pooled gene disruption, providing a robust population for studying ARFGAP2-dependent processes without clonal isolation. The polyclonal format ensures representation of a variety of editing events, ideal for functional genomics and pooled screening assays.
The HAP1 cell line is a near-haploid, adherent fibroblast-like derivative of chronic myeloid leukemia, widely employed for CRISPR knockout screens due to its single-copy genome. Its simplified karyotype facilitates unambiguous genotype?Cphenotype correlations and enhances the detection of subtle loss-of-function phenotypes. This background makes it particularly suitable for dissecting genes involved in membrane trafficking and organelle homeostasis.
ARFGAP2 encodes a GTPase-activating protein that specifically stimulates GTP hydrolysis on ARF1, converting active ARF1-GTP to inactive ARF1-GDP. This catalytic activity is essential for the timely disassembly of the COPI coat from vesicles, a pivotal event in retrograde trafficking from the Golgi to the ER and for maintaining Golgi integrity. ARFGAP2 is activated by ARF1-GTP and directly interacts with the COPI coatomer complex, Golgi SNAREs, and the KDEL receptor, integrating it into the ARF GTPase cycle. Loss of ARFGAP2 disrupts COPI-dependent vesicle budding, leading to a block in coat disassembly, reduced vesicle formation, and pronounced Golgi morphological defects.
In the HAP1 context, ARFGAP2 knockout provides a tractable system for investigating COPI-mediated trafficking and Golgi architecture. The near-haploid genome simplifies functional analysis, making this polyclonal population ideal for studying diseases linked to vesicular transport defects, such as Zellweger spectrum disorder and peroxisomal biogenesis disorders. The model captures a range of editing outcomes, reflecting population-level effects that mirror pooled genetic interaction screens.
Typical applications include immunofluorescence localization of Golgi markers such as Giantin and GM130, quantitative Western blot analysis of ARF1 and coatomer, and in vitro GTPase activity measurement. These polyclonal cells are also ideal for pooled drug sensitivity and genetic interaction screens aimed at dissecting COPI-dependent transport and Golgi function. For technical inquiries, please contact Ascent Research.
