The ARFIP2 Knouckout HT29 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal population of HT29 cells with targeted disruption of the ARFIP2 gene. This genetic perturbation creates a loss-of-function model for studying the scaffolding role of ARFIP2 in coordinating membrane trafficking and actin cytoskeleton dynamics. The polyclonal format represents a heterogeneous mixture of edited alleles, avoiding clonal selection and providing a population-level assessment of ARFIP2 deficiency. The knockout has been generated using CRISPR/Cas9 technology to introduce gene disruption, enabling researchers to interrogate ARFIP2 function without relying on transient knockdown approaches.
The HT29 host cell line is a well-established human colorectal adenocarcinoma model originating from a 44-year-old female patient. These epithelial cells harbor an APC mutation, a hallmark of colorectal cancer, and exhibit the capacity to differentiate under metabolic stress conditions such as glucose deprivation or butyrate treatment. HT29 cells are extensively utilized in intestinal epithelial biology and colorectal cancer research, including studies of tumor progression, metastasis, and therapeutic resistance. The tumorigenic background, combined with the APC mutation, provides a clinically relevant context for investigating molecular mechanisms driving colorectal malignancy.
ARFIP2 is an effector scaffold that links Arf and RAC1 GTPases, coupling Arf-mediated vesicle trafficking to RAC1-driven actin polymerization. It binds ARF1, ARF6, and RAC1, and associates with the WAVE regulatory complex and ARP2/3 nucleator. Upstream regulators include RAC1, ARF1, ARF6, and EGFR; downstream it promotes lamellipodia formation, membrane ruffling, and cell migration. By coordinating membrane trafficking with actin dynamics, ARFIP2 controls cell motility and invasion. Knockout impairs these processes, attenuating RAC1-dependent cytoskeletal reorganization and Arf-mediated endosomal transport.
In HT29 colorectal cancer cells, ARFIP2 knockout dissects the role of membrane trafficking?Cactin crosstalk in malignancy. The APC-mutant background increases relevance for metastasis studies, where migration and invasion are key. ARFIP2 loss attenuates RAC1 and ARF signaling downstream, impacting lamellipodial protrusion and adhesion. This model permits exploration of Arf-RAC1 pathway crosstalk with EGFR and how these networks affect epithelial plasticity and metastatic spread.
This knockout cell population supports diverse functional assays including wound healing and transwell migration/invasion, phalloidin-based actin visualization, co-immunoprecipitation for RAC1, western blotting for ARFIP2, and immunofluorescence. Cell adhesion and EGFR signaling analyses are also feasible. It enables research into colorectal cancer metastasis, Arf-RAC1 crosstalk, and actin dynamics. For technical details, contact Ascent Research.
