This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population targeting GRB10 (Growth factor receptor-bound protein 10) in A-549 human lung adenocarcinoma cells. The polyclonal population harbors heterogeneous loss-of-function mutations generated by CRISPR/Cas9-mediated gene disruption, providing a robust loss-of-function model that avoids clonal bias. This format enables functional interrogation of GRB10 across diverse genomic contexts within the same population.
The A-549 cell line, derived from lung adenocarcinoma of a 58-year-old Caucasian male, serves as a widely accepted model of human type II alveolar epithelial cells. Its established use in studying lung adenocarcinoma biology, including proliferation, metastasis, and drug response, makes it an appropriate host for exploring GRB10 function in a cancer-relevant setting. The knockout derivatives inherit these features, ensuring physiologically meaningful data.
GRB10 encodes an adaptor protein that directly interacts with activated INSR and IGF1R to inhibit receptor kinase activity and attenuate downstream signaling. This negative regulation suppresses the PI3K/AKT and MAPK/ERK cascades, impacting effectors such as IRS1, AKT, mTORC1, S6K1, and ERK1/2. GRB10 also interacts with NEDD4, RAF1, and MEK1, which modulate its function. Upstream, insulin, IGF-1, growth hormone, and the transcription factor EGR1 stimulate GRB10 expression, alongside epigenetic control via imprinting center methylation. Thus, GRB10 acts as a critical brake on growth and metabolic signaling pathways.
In A-549 cells, GRB10 knockout relieves inhibition on insulin/IGF-1 signaling, predicted to enhance PI3K/AKT and MAPK/ERK pathway activity. This can promote cell proliferation, survival, glucose uptake, and metabolic reprogramming??processes relevant to lung adenocarcinoma progression and therapy resistance. The model is therefore valuable for dissecting GRB10??s role in lung cancer and related signaling-driven diseases, including type 2 diabetes, obesity, and Silver-Russell syndrome. Its polyclonal nature further allows assessment of phenotypic variability in signaling outcomes.
This polyclonal knockout model supports diverse assays. Western blotting for phospho-AKT and phospho-ERK1/2 directly measures pathway activation, while RT-qPCR quantifies residual GRB10 transcripts. Cellular responses can be evaluated by MTT proliferation assays, Annexin V apoptosis assays, Transwell migration tests, and glucose uptake measurements. The model also enables genome-wide transcriptomic profiling by RNA-seq and epigenetic studies of GRB10 regulation. Researchers can apply this system to investigate lung adenocarcinoma biology, insulin/IGF-1 signaling, drug resistance mechanisms, and metabolic adaptations in cancer. For further information, please contact Ascent Research.
