The IFT88 Knockout DLD-1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal population of DLD-1 human colorectal adenocarcinoma cells carrying a targeted disruption of the IFT88 gene. This polyclonal knockout cell pool provides a physiologically relevant loss-of-function model for investigating the roles of intraflagellar transport and primary cilia in colorectal cancer signaling.
The DLD-1 cell line is a widely used model of colorectal adenocarcinoma, characterized by microsatellite instability high (MSI-H) and oncogenic mutations in KRAS (G13D) and APC. These genetic alterations drive aberrant Wnt and MAPK pathway activation, making DLD-1 a robust system for studying colorectal cancer progression, epithelial-mesenchymal transition (EMT), and drug resistance mechanisms.
IFT88 encodes a core component of the intraflagellar transport (IFT) complex B, essential for anterograde transport during primary cilium assembly and maintenance. IFT88 interacts with multiple IFT-B partners, including IFT52, IFT57, IFT74, IFT81, and IFT20, and cooperates with molecular motors such as KIF3A and DYNC2H1. Its expression is regulated by ciliogenic transcription factors RFX1, RFX3, and FOXJ1. Disruption of IFT88 abolishes ciliogenesis, thereby silencing cilia-dependent signal transduction. Key pathways affected include Hedgehog signaling, where loss of IFT88 prevents GLI1/2 activation downstream of SMO and PTCH1; Wnt/??-catenin signaling, where ciliary defects alter CTNNB1 stability and TCF/LEF-mediated transcription of targets like CCND1 and AXIN2; and TGF-?? signaling, where impaired ciliary function reduces phospho-SMAD2/3 levels downstream of TGFBR1.
In colorectal cancer, primary cilia have been implicated as signaling hubs that modulate pathways critical for tumor initiation and progression. The IFT88 knockout in DLD-1 cells provides a unique tool to dissect how ciliary disruption influences oncogenic KRAS and APC-driven phenotypes. By blocking ciliogenesis, this model permits examination of altered Hedgehog, Wnt, and TGF-?? pathway outputs, with measurable effects on cellular proliferation, epithelial-mesenchymal transition, and invasive behavior. Thus, it enables functional studies of ciliary signaling in a molecularly defined colorectal cancer background.
Typical applications include mechanistic studies of ciliary signaling in colorectal cancer, ciliopathy disease modeling, and evaluation of pathway-specific inhibitors for Hedgehog or Wnt signaling. The polyclonal IFT88 knockout cells are well-suited for immunofluorescence-based cilia visualization using markers such as acetylated tubulin and ARL13B, western blotting and RT-qPCR to confirm IFT88 loss, and functional assays including Gli-luciferase reporter activation, cell proliferation, wound healing, and transwell invasion. Downstream transcriptomic changes can be assessed via RNA-seq. For further information or to discuss custom project requirements, please contact Ascent Research.
