The DYNLT1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HT29 human colorectal adenocarcinoma cells, designed for disruption of DYNLT1. This gene encodes the cytoplasmic dynein light chain Tctex-type 1. The polyclonal format provides a heterogeneous pool of edited cells, avoiding clonal selection bottlenecks while enabling robust loss-of-function studies. CRISPR/Cas9-mediated gene disruption has been employed to generate this knockout model, suitable for investigating dynein-dependent processes in an epithelial cancer context.
HT29 is a widely used epithelial colorectal adenocarcinoma cell line with well-characterized mutations in APC and TP53, leading to constitutive Wnt pathway activation. It serves as a model for colon cancer biology, intestinal barrier function, and drug transport. HT29 cells form polarized monolayers and retain some differentiation features, making them ideal for studying mechanisms of cell polarity, adhesion, and migration in a tumor-relevant setting.
DYNLT1 protein is a subunit of the cytoplasmic dynein motor complex, mediating retrograde microtubule-based transport of vesicles, organelles, and protein complexes. It also exerts dynein-independent functions by binding to Fyn kinase and influencing its subcellular localization. DYNLT1 activity is regulated by phosphorylation via CDK1 and Aurora A kinases, and it interacts with dynein intermediate chain (DYNC1I1), dynein heavy chain (DYNC1H1), dynactin (p150Glued), NUDEL, and LIS1. In HT29 cells, DYNLT1 knockout disrupts mitotic spindle orientation, focal adhesion dynamics, and trafficking of E-cadherin and ??-catenin, potentially impairing Wnt/??-catenin signaling and altering cell migration and genomic stability.
Loss of DYNLT1 in the colorectal cancer context provides insights into how dynein-dependent transport contributes to malignant phenotypes. Impaired spindle assembly may lead to chromosome missegregation and genomic instability, while defective recycling of adhesion molecules can enhance invasive behavior. Since HT29 cells are often used in drug response studies, DYNLT1 knockout may also reveal roles of intracellular trafficking in chemosensitivity. The interplay with Wnt pathway components highlights the model’s utility for investigating colorectal carcinogenesis and metastasis.
Typical applications include immunofluorescence analysis of mitotic spindle morphology, cell cycle analysis by flow cytometry, wound healing and transwell invasion assays to measure migration and invasion, co-immunoprecipitation to assess dynein complex integrity, and western blotting for phosphorylated DYNLT1. Effects on Wnt target genes can be examined by qRT-PCR. Drug sensitivity screening can identify compounds whose efficacy is modulated by dynein function. For ordering, contact Ascent Research.
