The ID1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of human NCI-H1975 lung adenocarcinoma epithelial cells, engineered for targeted disruption of the ID1 gene. This polyclonal format provides a heterogeneous pool of edited cells, avoiding clonal selection artifacts and preserving genetic diversity for functional studies. The product enables robust loss-of-function analyses without the biases of single-cell cloning, making it suitable for population-level assessments of ID1-dependent phenotypes.
The parental NCI-H1975 cell line was derived from a lung adenocarcinoma of a nonsmoking female and harbors activating EGFR L858R and T790M mutations, with wild-type TP53. This genetic background establishes the cells as a clinically relevant model for EGFR-mutant non-small cell lung cancer (NSCLC), widely used to study acquired resistance to EGFR tyrosine kinase inhibitors (TKIs) and to evaluate next-generation therapeutic strategies.
ID1 functions as a dominant-negative inhibitor of basic helix-loop-helix (bHLH) transcription factors, including E2A (TCF3), HEB (TCF12), and E2-2 (TCF4). By heterodimerizing with these bHLH proteins, ID1 prevents their DNA binding and transcriptional activity, thereby promoting cell proliferation and survival while blocking differentiation. ID1 is transcriptionally activated by BMP2/BMP4 through SMAD1/5/8, TGF-beta1, EGF, HIF-1alpha, and other upstream regulators. It controls downstream effectors such as MMP2, MMP9, VEGF, Cyclin D1, Bcl-2, and p21, thereby integrating signals from the TGF-beta/BMP, PI3K/AKT, MAPK/ERK, and Wnt pathways to drive oncogenic processes.
In NCI-H1975 cells, EGFR L858R/T790M mutations drive persistent signaling that frequently upregulates ID1, contributing to epithelial-mesenchymal transition (EMT), invasion, and TKI resistance. Disruption of ID1 in these cells is expected to derepress bHLH targets, leading to increased expression of CDK inhibitors p21 and p16INK4a, reduced MMP-mediated matrix degradation, and impaired migratory capacity. Consequently, ID1 knockout may attenuate downstream AKT and ERK survival signals, enhance sensitivity to EGFR TKIs such as osimertinib, and reduce tumorigenic potential in vivo.
This model supports applications such as studying ID1??s role in EGFR-mutant NSCLC using proliferation (MTT/CCK-8), apoptosis (Annexin V), and migration/invasion (Transwell) assays. Colony formation and xenograft studies assess clonogenicity and tumor growth, while drug sensitivity assays evaluate synergy with EGFR inhibitors. Co-immunoprecipitation confirms disrupted ID1?CE2A complexes, and RNA-seq or western blotting with RT-qPCR characterizes downstream targets (MMP2, VEGF, p21). Additional uses include EMT and cancer stemness research, synthetic lethality screens, and exploring BMP/TGF-beta-mediated drug resistance. For additional technical information, please contact Ascent Research.
