This CRISPR/Cas9-edited polyclonal knockout cell population is derived from the A-549 human lung adenocarcinoma epithelial cell line and carries targeted disruption of the IFT43 gene. IFT43 encodes a core component of the intraflagellar transport complex A (IFT-A), which is essential for retrograde ciliary transport and ciliary maintenance. The polyclonal format yields a heterogeneous pool of IFT43 loss-of-function mutations, providing a robust, non-clonal model that captures diverse genetic knockout outcomes and minimizes artifacts associated with single-cell-derived lines.
The parental A-549 cell line was originally established from the lung adenocarcinoma tissue of a 58-year-old Caucasian male and serves as a well-characterized model system for lung adenocarcinoma biology, drug sensitivity profiling, and viral infection studies. These epithelial cells exhibit the capacity to assemble primary cilia under defined culture conditions, making them particularly amenable to investigations of ciliogenesis. Disruption of IFT43 is predicted to severely impair the assembly, maintenance, and signaling functions of the primary cilium in this cancer background.
Mechanistically, IFT43 is an integral IFT-A subunit that, together with IFT-B and the BBSome, drives retrograde ciliary transport. Its expression is regulated by ciliogenic transcription factors RFX3 and FOXJ1. IFT43 interacts with IFT-A components IFT121, IFT122, IFT139, IFT140, and IFT144; loss of IFT43 disrupts the complex and blocks retrograde trafficking. This impairs Hedgehog signaling by affecting Smoothened (SMO) and Gli transcription factors, and alters Wnt signaling through Dishevelled (DVL), AXIN, and beta-catenin, attenuating downstream transcriptional programs.
In A-549 lung adenocarcinoma cells, IFT43 knockout can modify proliferation, migration, and invasion phenotypes linked to ciliary signaling. This polyclonal model enables dissection of cilia-dependent pathways in cancer biology and may recapitulate molecular features of ciliopathies such as cranioectodermal dysplasia and short-rib thoracic dysplasia in an epithelial setting.
Applications include ciliogenesis assays, cilia length measurements (acetylated tubulin/ARL13B immunofluorescence), GLI-luciferase Hedgehog reporter studies, RNA-seq transcriptomics, and functional migration/proliferation assays. The cells are ideal for drug screening targeting ciliopathy or oncogenic ciliary signaling. Contact Ascent Research for further details and ordering information.
