The KRCC1 Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line, engineered to disrupt the KRCC1 gene. This pooled knockout model provides a heterogeneous cell population with targeted gene disruption, enabling robust loss-of-function studies in a physiologically relevant non-small cell lung cancer (NSCLC) background. By introducing CRISPR/Cas9-mediated gene disruption, the product allows researchers to dissect KRCC1-dependent signaling mechanisms without the constraints of clonal selection, capturing population-level biological variability.
The NCI-H1975 cell line is a widely utilized human lung adenocarcinoma model established from a female patient, endogenously expressing EGFR L858R and T790M mutations. These mutations drive constitutive MAPK/ERK signaling, conferring partial resistance to first-generation EGFR inhibitors. NCI-H1975 thus serves as a standard model for studying EGFR-targeted therapy resistance and evaluating next-generation inhibitors, with its epithelial NSCLC origin providing a clinically relevant context for oncogenic signaling studies.
KRCC1 encodes a chromatin-associated protein that directly interacts with MAPK1 (ERK2), a central effector kinase of the MAPK/ERK cascade. KRCC1 functions as a modulator of ERK2 activity, influencing the phosphorylation and transcriptional regulation of downstream targets such as cyclin D1, c-Fos, c-Jun, and the transcription factor ELK1. The MAPK/ERK pathway is activated by upstream signals including epidermal growth factor receptor (EGFR) and the RAS-RAF-MEK kinase module; KRCC1 operates downstream of MEK1/2 and forms a regulatory node at the ERK2 level. Through its interaction with chromatin remodeling proteins, KRCC1 may also contribute to local chromatin organization, thereby tuning the accessibility of ERK2-responsive gene loci.
Disruption of KRCC1 in NCI-H1975 cells perturbs the KRCC1-ERK2 interaction, likely altering ERK2 signaling dynamics and downstream transcriptional programs. In the EGFR-mutant background, this knockout model facilitates exploration of how KRCC1 loss rewires proliferation and apoptosis control in NSCLC, and whether it modulates sensitivity to EGFR-targeted agents such as osimertinib or erlotinib. It also enables analysis of ERK2-dependent gene expression changes, such as c-FOS and cyclin D1, thereby illuminating the interplay between chromatin-associated ERK2 regulation and drug resistance mechanisms.
Common applications include western blotting for phosphorylated and total ERK2, RT-qPCR quantification of ERK target genes, and cell proliferation (MTS) and apoptosis (Annexin V) assays. Co-immunoprecipitation can confirm disrupted KRCC1-ERK2 binding, while RNA-seq transcriptomics captures global expression changes. Drug sensitivity testing with osimertinib or erlotinib assesses functional impacts on EGFR inhibitor response, and the model can be used in MAPK pathway inhibitor screens. For further details and ordering information, please contact Ascent Research.
