The ARL6IP5 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma line. The product consists of a heterogeneous pool of cells collectively carrying loss-of-function disruptions in the ARL6IP5 gene, enabling robust interrogation of ARL6IP5-dependent phenotypes without clonal selection artifacts. This polyclonal format is suited for bulk assays measuring migration, proliferation, signaling changes, and stress responses.
The HT29 parental line originates from a primary colorectal adenocarcinoma of a 44-year-old Caucasian female and exhibits adherent epithelial morphology. Widely used in colon cancer biology and intestinal drug absorption studies, HT29 cells provide a well-characterized model for oncogenic signaling, metastasis, and therapeutic resistance. Their genetic and phenotypic stability supports reproducible experimental outcomes across diverse assay formats.
ARL6IP5 encodes a multi-pass membrane protein that modulates excitatory amino acid transporters EAAT1 (SLC1A3) and EAAT2 (SLC1A2), regulating glutamate uptake and protecting against oxidative stress. The protein participates in endosomal trafficking and is regulated by transcription factors SP1 and AP-1, as well as by oxidative stimuli and retinoic acid. Downstream, ARL6IP5 influences the MAPK/ERK cascade (through GRB2, SOS, RAS, RAF, MEK, ERK) and the PI3K/AKT pathway (PI3K, AKT, mTOR), affecting cell proliferation, survival, and migration. It also modulates apoptosis-related factors such as BCL2, BAX, CASP3, and the cell cycle inhibitor CDKN1A, and interacts with ARL6, EAATs, and the addicsin complex.
In HT29 cells, ARL6IP5 knockout is expected to disrupt glutamate homeostasis, increase sensitivity to oxidative stress, and enhance migration and proliferation through dysregulation of MAPK/ERK and PI3K/AKT pathways. This model enables investigation of how ARL6IP5 loss shifts the balance between cell death and survival in colorectal cancer, and its polyclonal nature better recapitulates tumor heterogeneity for drug response studies.
Researchers can employ these cells in glutamate uptake assays, Western blotting, RT-qPCR, MTT proliferation assays, Annexin V apoptosis detection, transwell migration assays, colony formation, ROS measurement, and co-immunoprecipitation to examine protein interactions. Applications span studies of colon cancer progression, oxidative stress signaling, drug resistance, and metastasis. For further information, please contact Ascent Research.
