The EFHD1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population featuring disruption of the EFHD1 gene in the NCI-H1975 human lung adenocarcinoma cell line. This loss-of-function model consists of a heterogeneous pool of edited cells, maintaining the host line??s genetic background while eliminating EFHD1 expression. The polyclonal format avoids the selective pressure of single-cell cloning, providing a more physiologically relevant system that mirrors tumor cell heterogeneity and is well-suited for functional gene studies.
The host cell line NCI-H1975 is a widely utilized non-small cell lung cancer model derived from a female patient. These epithelial cells harbor activating EGFR L858R/T790M mutations, conferring constitutive kinase activity and resistance to first-generation tyrosine kinase inhibitors. NCI-H1975 is extensively applied to study EGFR-driven signal transduction, tumor progression, and mechanisms of drug resistance in lung adenocarcinoma.
EFHD1 is a calcium-binding EF-hand protein that regulates actin cytoskeleton organization and cell adhesion. It interacts with cofilin to modulate actin polymerization and binds 14-3-3 proteins, linking calcium signals to cytoskeletal dynamics. EFHD1 is activated by intracellular calcium and functions downstream of EGFR signaling, hypoxia, and TGF-??. It controls apoptotic signaling through BCL2 family proteins and caspase-3/7 activation, and influences FAK-mediated focal adhesion turnover.
In NCI-H1975 cells harboring EGFR L858R/T790M mutations, EFHD1 knockout provides a system to investigate calcium-binding protein contributions to EGFR inhibitor resistance, cytoskeletal reorganization, and apoptosis. This model is instrumental for studying how actin dynamics affect cell survival and invasion, potentially uncovering strategies to overcome drug resistance.
Applications include Transwell migration/invasion, wound healing, and Annexin V apoptosis assays. Western blotting of EFHD1, cofilin, cleaved caspases, and BCL2 members, immunofluorescence of F-actin and vinculin, calcium imaging, and co-immunoprecipitation are representative techniques. Used for research on cancer cell migration, calcium signaling, apoptosis, EGFR inhibitor resistance, and cytoskeletal dynamics. For further information, contact Ascent Research.
