The IFT20 Knockout A-549 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population originating from the human A-549 lung adenocarcinoma cell line, with targeted disruption of the IFT20 gene. This product provides a loss-of-function model for studying the intraflagellar transport protein IFT20, an essential component of the IFT-B complex. The polyclonal format ensures a heterogeneous knockout pool, avoiding clonal selection and capturing diverse editing outcomes.
The parental A-549 cell line is a widely used model derived from a 58-year-old male with lung adenocarcinoma. These epithelial cells display features of type II pneumocytes, including surfactant production and tight junction formation, and are employed to study alveolar barrier function, adenocarcinoma biology, and drug response. The A-549 background enables investigation of ciliary functions in a cancer-relevant context, as these cells can be induced to form primary cilia.
IFT20 functions within the IFT-B complex alongside IFT52 and IFT88 to mediate anterograde ciliary transport. It additionally interacts with GMAP210 and VAMP7 for Golgi-to-cilium vesicle trafficking. Transcriptional regulation by RFX factors and activation by Hedgehog and TGF-?? signaling converge on IFT20. Downstream, IFT20 facilitates ciliary SMO accumulation and GLI transcription factor activation, including GLI1. In immune contexts, IFT20 recruits to the TCR complex, binding CD3 chains and ZAP70 to coordinate immune synapse formation and actin remodeling.
Knockout of IFT20 in A-549 cells impairs primary cilium assembly, enabling dissection of cilia-dependent processes in lung adenocarcinoma. Since primary cilia transduce Hedgehog signals, this model allows evaluation of how IFT20 loss alters SHH-driven GLI transcriptional programs and affects tumor cell behavior. The polyclonal population mirrors genetic heterogeneity found in tumors, enhancing relevance for drug screening and studies of cilia-mediated proliferation and migration.
These cells support a range of assays: immunofluorescence-based ciliogenesis analysis, GLI reporter gene assays for Hedgehog pathway activity, and Western blotting for IFT20 and associated proteins (e.g., IFT88, GMAP210). Wound healing assays can assess migration, while drug challenges explore cilia-targeting therapies. Additional applications include ciliopathy research and immune synapse studies using TCR pathway readouts. For technical details and orders, contact Ascent Research.
