The IFT27 Knockout HT29 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the gene encoding intraflagellar transport protein 27 homolog (IFT27) in the human colorectal adenocarcinoma HT29 cell line. This loss-of-function model, generated by CRISPR/Cas9-mediated gene disruption, provides a heterogeneous cell pool suitable for studying IFT27-dependent processes without clonal bias. It is specifically designed to investigate ciliogenesis, ciliary signaling, and their roles in colorectal cancer biology.
HT29 cells are a well-characterized human colorectal adenocarcinoma-derived epithelial cell line harboring mutations in TP53 and APC, while retaining wild-type KRAS and BRAF alleles. Their epithelial morphology and genetic background make them a widely used model for colorectal tumorigenesis, especially for studying chromosomal instability and dysregulated Wnt signaling. The HT29 line can be induced to form primary cilia under specific culture conditions, adding utility for cilia-related research despite its transformed state.
IFT27 encodes a small GTPase that is a core member of the IFT-B complex, essential for anterograde intraflagellar transport and ciliogenesis. Its activity is modulated by GTP/GDP exchange and the transcription factor FOXJ1. IFT27 collaborates with IFT-B partners IFT20 and IFT81 to transport key ciliary membrane proteins such as Arl13b and Smoothened. It also interfaces with the BBSome (e.g., BBS1, BBS4) for ciliary cargo trafficking. Loss of IFT27 abolishes primary cilium formation, silencing cilia-dependent Hedgehog signaling as evidenced by reduced GLI1 and GLI2 transcriptional output.
Ablation of IFT27 in HT29 cells prevents ciliogenesis even upon serum starvation, offering a clean model for assessing cilia-dependent signaling in a colorectal cancer context. This polyclonal knockout enables dissection of Hedgehog pathway contributions to tumor cell phenotypes amid APC and TP53 mutations. It also serves as a platform for studying ciliopathy-related mechanisms, including those underlying short-rib thoracic dysplasia, and for exploring interactions between ciliary and oncogenic signaling networks.
Typical experiments include immunofluorescence detection of ciliary markers (acetylated tubulin, Arl13b) to confirm cilia loss, RT-qPCR quantification of Hedgehog target genes (GLI1, PTCH1), and Western blotting for IFT27 expression. Functional assays such as MTT/WST-1 proliferation tests and soft agar colony formation can evaluate changes in cell growth. The polyclonal format also supports screening for small molecules that modulate ciliogenesis. For further details, contact Ascent Research.
