DYNLT3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited knockout cell population in which the DYNLT3 gene has been disrupted in the HT29 human colorectal adenocarcinoma cell line. As a loss-of-function model, these cells enable detailed investigation of the DYNLT3-encoded dynein light chain subunit and its roles in intracellular transport, mitotic progression, and cell signaling. The polyclonal nature of this knockout pool preserves population heterogeneity, avoiding the selection biases associated with single-cell cloning, and thereby more accurately reflects the diversity seen in tumor cell populations.
The HT29 cell line originates from a 44-year-old female Caucasian with colorectal adenocarcinoma and is extensively characterized as a model of intestinal epithelial cells. These cells harbor mutations commonly found in colorectal cancer, including constitutively active Wnt/??-catenin signaling due to APC mutation and defective p53 function, making them a physiologically relevant host for studying oncogenic processes and therapeutic responses.
DYNLT3 encodes a light chain subunit of the cytoplasmic dynein motor complex, which drives retrograde transport of vesicles, organelles, and protein complexes along microtubules. Within the dynein complex, DYNLT3 directly interacts with the dynein intermediate chain, dynein heavy chain, and the dynactin complex, and also associates with BICD2 and NUMA to regulate cargo motility and mitotic spindle assembly. The expression of DYNLT3 is transcriptionally regulated by Wnt/??-catenin/TCF, TGF-??/Smad pathways, and the transcription factor Sp1. Functionally, DYNLT3 influences downstream targets including p53 and components of the mitotic spindle apparatus, thereby integrating transport functions with cell cycle progression and apoptotic signaling.
In the HT29 colorectal cancer model, DYNLT3 knockout is anticipated to impair dynein-dependent processes critical for tumor cell physiology. Disruption of retrograde transport may lead to mislocalization of signaling molecules such as ??-catenin, potentially attenuating Wnt pathway output. Furthermore, defects in mitotic spindle organization can cause chromosome missegregation and genomic instability, hallmarks of colorectal carcinomas. This knockout model thus provides a rigorous platform to dissect the contributions of dynein light chain function to cancer cell proliferation, migration, and resistance to chemotherapeutics.
These polyclonal knockout cells are well-suited for a wide range of functional assays. Live-cell imaging can be used to track real-time organelle transport and endosomal trafficking. Mitotic index determination and flow cytometry-based cell cycle profiling allow assessment of mitotic defects. Western blotting and immunofluorescence enable confirmation of DYNLT3 knockout and analysis of downstream signaling components, while co-immunoprecipitation reveals alterations in dynein complex integrity. Transcriptomic approaches like RNA-seq can uncover global gene expression changes. The cells are also applicable to drug screening for dynein inhibitors and to studies of ciliopathies and neurodegenerative diseases. For additional information, please contact Ascent Research.
