The KIAA1217 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human NCI-H1975 lung adenocarcinoma line, carrying a targeted disruption of the KIAA1217 (DIAPH3) gene. This gene-edited pool provides a heterogeneous loss-of-function model for studying KIAA1217-dependent processes without single-cell cloning artifacts. The polyclonal format, generated via CRISPR/Cas9-mediated gene disruption, allows assessment of collective knockout effects on cytoskeletal dynamics and cancer cell behavior.
The NCI-H1975 cell line is an epithelial non-small cell lung adenocarcinoma model harboring EGFR L858R and T790M mutations, rendering it constitutively active and resistant to first-generation tyrosine kinase inhibitors. Widely used in oncology, this cell line is particularly suited for investigating mechanisms of tumor progression, drug resistance, and metastasis in EGFR-mutant lung cancer.
KIAA1217 encodes DIAPH3, a formin protein that nucleates unbranched actin filaments, playing a critical role in actin cytoskeleton organization, cell migration, and cytokinesis. It is activated downstream of RhoA and Rac1, phosphorylated by ROCK1, and modulated by phosphoinositides. DIAPH3 interacts with profilin and G-actin to facilitate filament elongation, and with Src and FAK at focal adhesions. Its activity drives actin polymerization and MRTF/SRF transcriptional responses, positioning it as a key node in the RhoA??ROCK1??DIAPH3??actin polymerization??cell migration pathway.
In the NCI-H1975 background, disruption of KIAA1217 is expected to impair actin-dependent processes essential for cancer cell dissemination, including migration and invasion. By eliminating DIAPH3-mediated actin nucleation, this polyclonal knockout model enables investigation of tumor cell behavior under attenuated metastatic conditions while maintaining EGFR-driven oncogenic signaling. It provides a platform to dissect the interplay between oncogenic pathways and cytoskeletal effectors in lung adenocarcinoma.
These knockout cells are suitable for a variety of experimental applications, including transwell migration/invasion assays, wound healing studies, and phalloidin staining to visualize actin defects. Western blotting and immunofluorescence can confirm DIAPH3 depletion and assess downstream targets such as FAK and MRTF/SRF. RNA-seq may reveal transcriptional alterations linked to actin dynamics, and drug screening can identify compounds targeting metastasis. Additionally, the model is relevant for studying DIAPH3-related hearing loss (DFNA1). For further information, please contact Ascent Research.
