The DNAAF2 Knockout NCI-H1975 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal cell population derived from the NCI-H1975 human lung adenocarcinoma cell line, in which the DNAAF2 gene has been disrupted to abolish its functional expression. This polyclonal knockout format provides a heterogeneous pool of cells harboring gene-disrupted alleles, enabling robust loss-of-function analysis without clonal selection artefacts. The cells serve as a powerful tool for investigating DNAAF2-dependent biological processes and for screening therapeutic strategies targeting ciliary dysfunction.
The NCI-H1975 cell line was originally established from the pleural effusion of a female non-smoker with lung adenocarcinoma, and it exhibits epithelial morphology. This line carries activating mutations in the epidermal growth factor receptor (EGFR) gene, specifically the L858R point mutation and the T790M gatekeeper mutation, which render it dependent on EGFR signaling and make it a widely used model for non-small cell lung cancer (NSCLC) research, particularly for studying acquired resistance to first-generation tyrosine kinase inhibitors.
DNAAF2 encodes a cytoplasmic co-chaperone protein that is essential for the preassembly of dynein arm complexes in motile cilia. It functions by interacting with molecular chaperones such as HSP70 and HSP90, as well as with dynein intermediate chains and the DNAAF1 protein, to facilitate the correct folding and assembly of outer and inner dynein arm subunits. The expression of DNAAF2 is transcriptionally regulated by the FOXJ1 and RFX family transcription factors, which are master regulators of ciliogenesis, and is modulated by NOTCH signaling. Disruption of DNAAF2 prevents proper dynein arm formation, leading to defective ciliary axonemal assembly and impaired ciliary beating.
In the context of NCI-H1975 adenocarcinoma cells, loss of DNAAF2 function provides a unique model to study the intersection of oncogenic EGFR signaling and ciliary biology. Lung adenocarcinoma cells may retain rudimentary cilia, and DNAAF2 knockout can abrogate residual ciliary motility, potentially affecting processes such as cell migration and response to mechanical stimuli. This model enables dissection of how ciliary dysfunction contributes to tumor cell behavior and may reveal synthetic vulnerabilities in EGFR-mutant tumors with compromised mucociliary clearance.
This polyclonal knockout product is well-suited for a broad range of experimental applications, including functional studies of DNAAF2 in lung cancer, investigation of ciliary motility mechanisms, and utilization as a model for primary ciliary dyskinesia (PCD) and Kartagener syndrome. Typical assays include western blotting and RT-qPCR for confirmation of DNAAF2 disruption, immunofluorescence staining for ciliary markers, ciliary beat frequency analysis, RNA sequencing for transcriptomic profiling, and cell migration assays to assess functional consequences. For further information, please contact Ascent Research.
