The KHDRBS1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma epithelial cell line, featuring targeted disruption of the KHDRBS1 gene. This loss-of-function model eliminates KHDRBS1 (Sam68) protein expression, enabling systematic investigation of its functions in alternative splicing and oncogenic signaling. The polyclonal format provides a heterogeneous pool of edited cells, broadly reflecting the diversity of CRISPR-mediated mutations, and is ideal for functional genomics studies where clonal variation is not required.
The parental NCI-H1975 cell line originates from a female patient with non-small cell lung adenocarcinoma and carries EGFR L858R and T790M mutations, which confer sensitivity and acquired resistance to first-generation tyrosine kinase inhibitors. These cells are extensively characterized as a model of EGFR-driven lung cancer, exhibiting constitutive activation of downstream MAPK/ERK and PI3K/AKT pathways, and are widely used to study oncogenic signaling, drug resistance, and metastasis. The NCI-H1975 background thus provides a clinically relevant platform for investigating crosstalk between EGFR signaling and RNA-binding proteins.
KHDRBS1 functions as a signal transduction adaptor and RNA-binding protein that bridges tyrosine kinase signals to RNA metabolism. Upon EGFR stimulation, Src family kinases activate KHDRBS1, which then regulates alternative splicing of proliferation and apoptosis regulators, including CD44 and Bcl-x, and promotes Cyclin D1 expression at the transcriptional level. It interacts with key signaling molecules such as Src, Grb2, PLC??1, and PI3K, integrating input from the MAPK/ERK and PI3K/AKT cascades to control cell cycle progression, survival, and motility.
In the NCI-H1975 cell system, knockout of KHDRBS1 disrupts the link between EGFR-driven Src kinase activity and the post-transcriptional regulatory network that supports lung adenocarcinoma cell proliferation, migration, and drug resistance. By eliminating KHDRBS1, researchers can dissect how EGFR signaling rewires RNA processing to sustain tumorigenic phenotypes in an EGFR-mutant background. This model is particularly valuable for evaluating the contribution of alternative splicing changes to resistance against EGFR tyrosine kinase inhibitors and for identifying splicing-dependent vulnerabilities that may serve as therapeutic targets in non-small cell lung cancer.
These KHDRBS1 knockout cells are suited for a wide range of functional studies, including RNA-seq and splicing-sensitive RT-PCR to profile KHDRBS1-dependent splicing events, Western blotting and RT-qPCR to validate target gene expression changes, and phenotypic assays such as colony formation, wound healing, and transwell migration to assess proliferation and motility. Drug sensitivity and apoptosis assays can explore how KHDRBS1 loss alters response to EGFR inhibitors, while co-immunoprecipitation enables mapping of altered protein?Cprotein interactions within the Src and PI3K/AKT signaling axes. For further technical information or assistance with experimental design, please contact Ascent Research.
