The ARL15 Knouckout HT29 Polyclonal Cells product comprises a heterogeneous population of HT29 cells engineered via CRISPR/Cas9-mediated disruption of the ARL15 locus. This polyclonal knockout model provides a versatile tool for investigating ARL15-dependent functions without the selection of a single clonal isolate. The pooled format preserves genetic diversity within the cell population, approximating a more physiologically relevant system. The gene-editing approach introduces loss-of-function mutations across a spectrum of alleles, resulting in a mixed cell pool with variable but collectively significant reduction in ARL15 protein activity. This product is designed for researchers requiring robust gene perturbation without clonal bias.
The HT29 parent cell line is a well-characterized human colorectal adenocarcinoma line derived from a female patient. These epithelial cells are widely employed in cancer biology and drug discovery due to their reproducible growth characteristics, defined genetic background, and relevance to colorectal tumorigenesis. HT29 cells harbor mutations in genes such as APC and p53, and exhibit deregulated signaling pathways, including Wnt and PI3K/AKT. Notably, HT29 cells display metabolic features such as aerobic glycolysis and altered lipid metabolism, making them an appropriate host for studying the intersection of oncogenic signaling and metabolic pathways.
ARL15 encodes a small GTPase of the ARF-like family that functions as a molecular switch in insulin-regulated metabolic processes. Upon insulin stimulation, ARL15 is activated downstream of the insulin receptor, IRS1, PI3K, and AKT signaling cascade, and operates in concert with ARF guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs). In its active GTP-bound state, ARL15 promotes GLUT4 translocation to the plasma membrane, enhancing glucose uptake. Additionally, ARL15 modulates the AMPK pathway and influences adipogenesis by regulating the expression of lipogenic transcription factors such as SREBP1c and its target FASN. Thus, ARL15 coordinates both glucose and lipid homeostasis.
In the HT29 colorectal cancer context, disruption of ARL15 offers unique insights into how malignant cells rewire metabolic pathways to support proliferation and survival. Colorectal cancers often exhibit insulin resistance and altered adipogenic programs, and HT29 cells serve as a model to dissect these processes. ARL15 knockout impairs insulin-mediated glucose uptake and lipogenic gene expression. This cell model enables the study of metabolic vulnerabilities in colorectal adenocarcinoma and the exploration of ARL15 as a node linking oncogenic and metabolic signals.
Researchers can employ these polyclonal knockout cells in a variety of experimental settings, including insulin resistance modeling, candidate drug evaluation for type 2 diabetes, and metabolic reprogramming investigations. Compatible assays include glucose uptake measurements, Western blotting for phosphorylated AKT and AMPK, RT-qPCR for GLUT4 and SREBP1c transcripts, Oil Red O lipid accumulation staining, and cell proliferation and migration assays. The cells are suitable for both mechanistic studies and high-throughput screening campaigns. For further details or to discuss custom applications, please contact Ascent Research.
