The DYNC2LI1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line. This product provides a loss-of-function model for the DYNC2LI1 gene, encoding a cytoplasmic dynein 2 light intermediate chain essential for retrograde intraflagellar transport (IFT) and ciliogenesis. The polyclonal format comprises a heterogeneous pool of edited cells, facilitating functional studies without single-cell cloning. By disrupting DYNC2LI1, researchers can investigate its role in ciliary maintenance and Hedgehog signaling within an epithelial context.
HT29 cells are a well-established model of human colorectal adenocarcinoma, widely employed for studies of intestinal barrier function, drug transport, and cancer biology. This epithelial line can form primary cilia under appropriate conditions, providing a physiologically relevant platform for examining ciliary mechanisms. The HT29 background thus offers a suitable context for investigating DYNC2LI1-dependent processes.
The DYNC2LI1 gene product is a subunit of the retrograde IFT dynein motor, essential for transporting cargo from the ciliary tip to the cell body. This retrograde transport is critical for cilium maintenance and Hedgehog signal transduction. Upstream, SHH ligand and FOXJ1 regulate ciliary gene expression. DYNC2LI1 interacts with DYNC2H1 and IFT proteins like IFT20 and IFT88. Disruption impairs retrograde transport, attenuating Hedgehog target gene activation (GLI1, GLI2, PTCH1). In the canonical pathway, SHH binding to PTCH1 relieves SMO inhibition, leading to GLI-mediated transcription; DYNC2LI1-dependent IFT ensures proper signal modulation.
In the HT29 cellular context, knockout of DYNC2LI1 is expected to compromise primary cilium formation and weaken Hedgehog pathway responsiveness. This combination enables the study of ciliary dysfunction in a cancer-relevant epithelial model. Since HT29 cells are derived from colorectal adenocarcinoma, the model may help elucidate how ciliary defects intersect with oncogenic signaling. The polyclonal knockout population allows assessment of bulk cellular phenotypes, such as ciliary frequency and Hedgehog-dependent transcription, without clonal selection biases. This approach is particularly useful for pharmacological screens aimed at restoring ciliogenesis or modulating the Hedgehog pathway in a pathophysiologically relevant setting.
Typical applications of these DYNC2LI1 knockout HT29 polyclonal cells include mechanistic studies of ciliogenesis, analysis of IFT dynamics, and investigation of Hedgehog signaling using techniques such as immunofluorescence for ciliary markers (acetylated ???tubulin, ARL13B), RT?qPCR for GLI1 and PTCH1 expression, and western blotting for dynein subunits. The cells are also suited for SMO agonist?induced reporter assays and high?content imaging to quantify ciliary length and frequency. Additionally, they provide a platform for identifying small molecules that can rescue ciliopathy?associated phenotypes. For further details or assistance, please contact Ascent Research.
