The KIDINS220 Knockout A-549 Polyclonal Cells product provides a CRISPR/Cas9-mediated gene-disrupted polyclonal population of A-549 human lung adenocarcinoma epithelial cells, targeting the scaffold protein KIDINS220. This loss-of-function model is designed for researchers investigating KIDINS220-dependent signaling mechanisms in non-small cell lung cancer and related cellular processes.
A-549 cells are a widely used adherent epithelial model derived from a 58-year-old male patient with lung adenocarcinoma, harboring a KRAS G12S activating mutation and wild-type TP53 status. These cells exhibit robust growth characteristics and are well-established for studying oncogenic signaling, drug response, and metastatic behaviour in vitro.
KIDINS220 (Kinase D-interacting substrate of 220 kDa) is a multi-domain scaffold that couples activated neurotrophin receptors (TrkA, TrkB) and growth factor receptors such as EGFR to downstream signaling cascades. Through direct interactions with kinases including Src and Fyn, KIDINS220 coordinates the assembly of signaling complexes that drive ERK1/2 and AKT phosphorylation via the Shc-Grb2-SOS-Ras-Raf-MEK pathway and PI3K-dependent activation, respectively. Additionally, KIDINS220 links to PLC??1-mediated calcium flux and associates with the p75 neurotrophin receptor and adaptors CrkL/CrkII to modulate NF-??B, CREB, and c-Jun transcriptional programs, thereby regulating cell survival, differentiation, and migration.
In the A-549 lung cancer context, KIDINS220 is implicated in sustaining proliferative and invasive signals. The A-549 cell line’s KRAS G12S mutation drives constitutive activation of the MAPK pathway, and scaffold proteins such as KIDINS220 can further amplify oncogenic signaling by clustering effectors. CRISPR/Cas9-mediated disruption of KIDINS220 in this polyclonal pool is anticipated to impair ERK and AKT activation downstream of KRAS and receptor tyrosine kinases, leading to reduced tumor cell growth, attenuated migration, and enhanced sensitivity to apoptotic stimuli. Consequently, this model enables dissection of KIDINS220’s contribution to malignant phenotypes and may reveal synthetic vulnerabilities.
Typical applications include Western blotting for phospho-ERK and phospho-AKT to assess signaling output, MTT proliferation and Transwell migration assays to quantify functional consequences, co-immunoprecipitation to map interactomes, and drug sensitivity testing for pathway inhibitors. The polyclonal format allows rapid expansion and experimental versatility while preserving heterogeneous knockout efficiencies across the population, facilitating pooled functional genomics and high-throughput screening applications. For additional details or technical support, please contact Ascent Research.
