The IFT25 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, designed to disrupt the IFT25 gene. This heterogeneous pool of cells with targeted gene disruption provides a versatile loss-of-function model for studying ciliary function and signaling in an epithelial tumor context.
The HT29 cell line was originally isolated from a primary colon adenocarcinoma of a 44-year-old Caucasian female. These cells exhibit absorptive and mucin-producing properties characteristic of intestinal epithelium and are widely employed in colorectal cancer research to investigate differentiation, signal transduction, and tumorigenic processes.
IFT25 is an essential subunit of intraflagellar transport (IFT) complex B, which moves cargo in the anterograde direction along ciliary axonemes. It interacts with multiple IFT complex B partners, including IFT27, IFT70, IFT52, IFT46, and IFT20, and cooperates with the kinesin-2 motor and cytoplasmic dynein 2. IFT25 disruption impairs ciliogenesis and severely reduces ciliary localization of key signaling molecules such as Smoothened (SMO) and polycystin-2 (PKD2). This, in turn, attenuates Hedgehog pathway activity by preventing proper processing and activation of Gli transcription factors (Gli1, Gli2, Gli3). Upstream, IFT25 expression is regulated by RFX transcription factors (RFX2, RFX3), and its function integrates with pathway components including Patched (PTCH1) and SUFU.
Loss of IFT25 in HT29 cells offers a unique opportunity to examine the role of primary cilia in colorectal cancer. Given that aberrant Hedgehog and Wnt signaling are frequently implicated in colorectal tumor progression, this knockout model enables dissection of how ciliary defects influence SMO trafficking, Gli-mediated transcription, and downstream cellular responses such as proliferation, migration, and drug sensitivity.
Researchers can apply this polyclonal knockout cell population in a range of assays, including immunofluorescence staining for ciliary markers (acetylated tubulin, Arl13b) to quantify cilia length and morphology, western blotting for IFT25 and Gli proteins, and RT-qPCR for Hedgehog target genes (Gli1, PTCH1). Functional studies??such as cell proliferation, scratch wound migration, colony formation, and drug sensitivity testing with Hedgehog inhibitors??are readily performed to assess the impact of IFT25 loss on tumorigenic properties. For further information about this product or to discuss custom gene-edited cell solutions, please contact Ascent Research.
