The HOOK3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colon adenocarcinoma line. This product features targeted disruption of the HOOK3 gene, encoding a microtubule-tethering adaptor critical for organelle positioning and intracellular trafficking. The polyclonal format provides a heterogeneous pool of edited cells, facilitating robust loss-of-function studies without single-cell cloning. The resulting HOOK3 loss-of-function model enables dissection of autophagy, endocytic transport, and microtubule-dependent organelle dynamics.
HT29 cells, isolated from a primary colon adenocarcinoma of a 44-year-old female, are a well-characterized epithelial model of colorectal cancer. They exhibit typical epithelial morphology, mucus secretion, and can differentiate into enterocyte-like cells. As a widely used intestinal epithelial line, HT29 is ideal for investigating cancer cell signaling, polarity, metabolism, and drug responses, providing a relevant host for gene knockout studies.
HOOK3 functions as a microtubule-tethering protein that bridges endosomes, lysosomes, and the Golgi apparatus to the microtubule network, regulating their perinuclear positioning and intracellular trafficking. It forms the FTS/HOOK/FHIP complex with FTS (AKTIP) and FHIP, linking cargoes to the dynein-dynactin motor. Upstream, Rab5 and Rab7 GTPases and autophagy-inducing stimuli regulate HOOK3 activity. Downstream, HOOK3 promotes LC3 lipidation and autophagosome-lysosome fusion, and maintains endosomal and Golgi morphology, interacting with RILP for lysosomal trafficking. Thus, HOOK3 integrates endocytic and autophagic signals to orchestrate organelle dynamics.
In HT29 colorectal adenocarcinoma cells, HOOK3 knockout disrupts critical trafficking processes, impairing autophagy and endosomal positioning, which may alter metabolic adaptation, stress responses, and cellular homeostasis. This perturbation can affect cell polarity, migration, and secretion, making the model valuable for studying how organelle trafficking influences colorectal cancer progression and potential vulnerabilities. It also enables exploration of HOOK3??s putative link to Charcot-Marie-Tooth neuropathy and cellular trafficking disorders in an epithelial cancer background.
This polyclonal knockout cell population is ideal for dissecting autophagy and endocytic trafficking using western blotting for LC3 and p62, immunofluorescence for LAMP1 and Golgi markers, and autophagy flux assays with chloroquine. Endocytosis, migration, invasion, and proliferation assays combine with functional genomics approaches like RNA-seq to profile transcriptomic changes. Applications extend to drug screening for autophagy modulators and studies of microtubule-dependent events in cancer metabolism. For further details, please contact Ascent Research.
