The IFT140 Knockout HT29 Polyclonal Cells product is a heterogeneous population of HT29 colorectal adenocarcinoma cells engineered by CRISPR/Cas9-mediated disruption of the IFT140 gene. This polyclonal knockout pool provides a loss-of-function model for investigating intraflagellar transport complex A (IFT-A) function and ciliary signaling dynamics. As a bulk-edited population, it avoids clonal selection biases, enabling robust functional analyses without single-cell cloning artifacts.
The parental HT29 cell line is a human colorectal adenocarcinoma model established from a primary tumor and widely used to study intestinal epithelial differentiation and colorectal cancer. These cells display features of absorptive enterocytes and can form primary cilia under serum-deprived conditions, making them suitable for ciliary investigations. Their capacity for enterocytic differentiation allows examination of ciliary roles in polarized epithelial function and oncogenic transformation.
IFT140 encodes a core subunit of the IFT-A complex, which mediates retrograde ciliary trafficking essential for ciliogenesis and Hedgehog signal transduction. IFT140 interacts with other IFT-A components such as IFT122 and IFT144, the cargo adaptor TULP3, and dynein-2 motor subunits DYNC2H1 and DYNC2LI1. Transcription factors RFX3 and FOXJ1 regulate IFT140 expression upstream. Disruption of IFT140 impairs the retrograde transport of signaling receptors, including Smoothened (SMO) and Patched1 (PTCH1), leading to attenuated Hedgehog pathway activity with reduced GLI1 and GLI2 transcriptional output and altered ??-catenin?Cmediated non-canonical Wnt signaling.
In the HT29 colorectal adenocarcinoma background, IFT140 knockout creates a unique model to dissect the interplay between ciliary signaling and intestinal epithelial homeostasis. Hedgehog pathway components like SMO and GLI1 influence differentiation and proliferation pathways frequently dysregulated in colorectal cancer. This system enables study of how loss of primary cilia affects tumor cell behavior, including migration, apoptosis, and drug sensitivity, and facilitates investigation of crosstalk between Hedgehog and Wnt signaling in a cancer-relevant context.
Key applications include investigating primary cilia function in colorectal cancer progression, screening for modulators of ciliopathy-associated pathways, and analyzing functional consequences of IFT140 loss. Representative assays include immunofluorescence staining for ciliary markers (acetylated ??-tubulin, ARL13B), Gli-luciferase reporter assays for Hedgehog pathway activity, wound-healing migration assays, annexin V apoptosis assays, and drug sensitivity profiling with agents such as vismodegib. For further technical details and ordering information, please contact Ascent Research.
