DYNC2LI1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population carrying targeted disruptions in the DYNC2LI1 gene within the NCI-H1975 human lung adenocarcinoma cell line. This loss-of-function model eliminates the light intermediate chain of cytoplasmic dynein-2, enabling functional studies of retrograde intraflagellar transport (IFT) and primary cilia biology. The polyclonal format preserves genetic heterogeneity and is suited for population-level assays, avoiding clonal selection artifacts.
The NCI-H1975 host line originates from a lung adenocarcinoma of a 62-year-old female nonsmoker and harbors two activating mutations: EGFR L858R and PIK3CA G118D. These oncogenic drivers activate MAPK and PI3K/AKT signaling, respectively, making the line a principal model for EGFR-targeted therapy resistance and PI3K pathway investigation in non-small cell lung cancer (NSCLC). Its adherent epithelial morphology facilitates standard cell culture and imaging workflows.
DYNC2LI1 encodes a subunit of the dynein-2 complex, essential for retrograde IFT within primary cilia. The dynein-2 motor, which includes DYNC2H1, WDR34, WDR60, TCTEX1D2, and DYNLT1, collaborates with IFT-A and IFT-B particles to transport cargo from the ciliary tip to the cell body. DYNC2LI1 expression is governed by RFX transcription factors and FOXJ1. Loss of functional DYNC2LI1 blocks retrograde trafficking, preventing activation of GLI transcription factors and suppressing Hedgehog target genes such as PTCH1, HHIP, and CCND1. Consequently, DYNC2LI1 knockout disrupts ciliogenesis and attenuates Hedgehog pathway output.
In the context of NCI-H1975 cells, primary cilia and Hedgehog signaling may intersect with oncogenic EGFR and PI3K pathways, potentially influencing tumor progression and therapy response. This knockout model permits dissection of cilia-dependent signaling in an EGFR-mutant background, enabling assessment of ciliary contributions to proliferation, migration, and tyrosine kinase inhibitor sensitivity. The concurrent PIK3CA mutation further allows exploration of PI3K/AKT crosstalk with IFT-dependent Hedgehog regulation.
This polyclonal knockout product supports diverse applications such as studying cilia biology in lung adenocarcinoma, Hedgehog pathway mechanistic investigations, ciliopathy modeling, and screening of SMO inhibitors (e.g., vismodegib). Representative assays include western blotting for DYNC2LI1, immunofluorescence for ARL13B and acetylated tubulin, RT-qPCR for GLI1 and PTCH1, cell cycle and migration assays, drug sensitivity profiling, RNA-seq, and flow cytometry. For additional technical details and ordering information, please contact Ascent Research.
