The CCDC91 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population with disruption of the CCDC91 gene in NCI-H1975 lung adenocarcinoma cells. This loss-of-function model facilitates investigation of CCDC91 roles in centriolar satellite biology, primary cilium assembly, and related signaling. The polyclonal format provides a heterogeneous pool of edited cells, avoiding clonal selection biases.
NCI-H1975 is an adherent epithelial line from pleural effusion of a 63-year-old female with non-small cell lung adenocarcinoma. It harbors EGFR L858R and T790M mutations, representing a key model of acquired resistance to EGFR tyrosine kinase inhibitors. The aneuploid line captures tumor heterogeneity and is widely employed to study drug resistance mechanisms, oncogenic signaling, and epithelial-mesenchymal transition in NSCLC.
CCDC91 encodes a centriolar satellite protein localizing to pericentriolar matrix, essential for primary cilium formation through regulation of microtubule organization and satellite integrity. It interacts with satellite components PCM1, CEP290, CEP131, SSX2IP, and OFD1, and recruits ciliary assembly factors like IFT88, BBS4, and ARL13B. Functioning downstream of the CP110-CEP97 complex and RFX transcription factors, CCDC91 is upstream of ciliogenesis-dependent Hedgehog and Wnt signaling. Its disruption attenuates Hedgehog pathway activation (reducing SMO, SUFU, and GLI1/3) and diminishes Wnt/??-catenin signaling (affecting AXIN2, DVL, and ??-catenin stability), leading to downregulation of downstream effectors MYC and Cyclin D1.
In NCI-H1975 cells driven by mutant EGFR, loss of CCDC91 and impaired ciliogenesis offer a system to probe interplay between EGFR, Hedgehog, and Wnt pathways. Because primary cilia can modulate receptor tyrosine kinase signaling and cell cycle entry, CCDC91 knockout may reveal vulnerabilities in EGFR-mutant NSCLC, especially under therapeutic pressure. This model enables dissection of how centriolar satellite dysfunction influences tumor cell proliferation, migration, and survival, and may uncover synthetic lethal interactions or targets for overcoming resistance.
These polyclonal knockout cells suit serum starvation-induced ciliogenesis assays, immunofluorescence for acetylated ??-tubulin/ARL13B, Western blotting for GLI proteins and EGFR signaling, and RT-qPCR for ciliary genes. Functional analyses include scratch wound migration, MTT viability, and co-immunoprecipitation of satellite complexes. They also allow assessment of Hedgehog/Wnt pathway responses during EGFR inhibitor treatment. For further details, contact Ascent Research.
