The IGF2BP1 Knockout NCI-H1975 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal population derived from human lung adenocarcinoma NCI-H1975 cells, with targeted disruption of the IGF2BP1 gene. This loss-of-function model enables examination of IGF2BP1-dependent post-transcriptional control in non-small cell lung cancer (NSCLC). The polyclonal format retains genetic heterogeneity while effectively eliminating target gene expression, making it suitable for pooled screens and population-level biochemical analyses. The cells are provided as a ready-to-use knockout pool, allowing direct assessment of IGF2BP1 ablation on mRNA stability and oncogenic signaling.
NCI-H1975 is a lung adenocarcinoma cell line with EGFR L858R and T790M mutations, widely used to investigate EGFR TKI resistance, particularly to osimertinib. This cell line is an established in vitro model for studying acquired resistance to third-generation EGFR inhibitors and for exploring signaling adaptations that bypass targeted therapy. Its EGFR-mutant background offers a clinically relevant context for evaluating the impact of IGF2BP1 knockout on tumor cell behavior and drug sensitivity.
IGF2BP1 is an oncofetal RNA-binding protein that recognizes m6A-modified mRNAs, enhancing their stability and translation. It targets MYC, KRAS, CTNNB1, and CD44 transcripts, promoting proliferative and stemness pathways. IGF2BP1 acts downstream of the ??-catenin/TCF transcription complex and MYC, and is regulated by microRNAs miR-98 and miR-1275. It interacts with translation initiation factors eIF4E, eIF4G, PABPC1, and the RNA-binding proteins HuR and YBX1 to facilitate efficient translation. Through these interactions, IGF2BP1 sustains Wnt/??-catenin, mTOR, and MAPK/ERK signaling, supporting tumorigenesis.
In NCI-H1975 cells, IGF2BP1 reinforces expression of MYC and KRAS, which are critical for EGFR-mutant NSCLC survival and TKI resistance. Knockout of IGF2BP1 disrupts m6A-dependent stabilization of these transcripts, reducing oncogenic protein levels and attenuating downstream effector pathways. This model enables dissection of post-transcriptional control intersecting with EGFR-driven signaling and characterization of clonal heterogeneity in drug resistance. The polyclonal knockout population is ideal for identifying synthetic lethal interactions that may sensitize tumors to targeted therapies.
Applications include proliferation assays (MTT, CellTiter-Glo), colony formation, and migration/invasion assays to assess tumorigenic properties. RNA immunoprecipitation and transcriptome-wide RNA-seq reveal altered mRNA target profiles and m6A dynamics. Drug sensitivity profiling with osimertinib evaluates therapeutic response modulation. The cells also support validation of RNA-based therapeutic targets and studies of EGFR TKI resistance mechanisms. For additional technical information, please contact Ascent Research.
