The CD44 Knockout NCI-H1299 Polyclonal Cells consist of a heterogeneous population of NCI-H1299 non-small cell lung carcinoma cells in which the CD44 gene has been disrupted by CRISPR/Cas9-mediated genome editing. This polyclonal knockout pool provides a loss-of-function model for studying CD44-dependent processes without the selective pressure of single-cell cloning.
NCI-H1299 is a human NSCLC cell line derived from a lymph node metastasis of a lung adenocarcinoma. The line is homozygous for TP53 deletion, rendering it p53-deficient and particularly useful for investigating p53-independent oncogenic mechanisms. NCI-H1299 cells display prominent migratory and invasive behavior, making them a standard model for NSCLC metastasis and drug resistance research.
CD44 encodes a transmembrane glycoprotein that functions as the major receptor for hyaluronan (HA). HA?CCD44 binding recruits ERM adaptor proteins and Src family kinases, triggering PI3K/AKT and MAPK/ERK signaling cascades. Activated AKT and ERK1/2 promote proliferation and survival by upregulating Cyclin D1, c-Myc, and MMP-9. CD44 also engages the NF-??B pathway via IKK activation and potentiates Wnt/??-catenin signaling through ??-catenin nuclear translocation, leading to expression of EMT transcription factors Snail and Slug. These pathways are modulated by upstream regulators such as TGF-??, EGF, and NF-??B, creating a network that controls adhesion, migration, and invasion.
In NCI-H1299 cells, CD44 is frequently overexpressed and contributes to aggressive metastatic traits and chemoresistance. Disruption of CD44 in this p53-null background allows researchers to dissect its role in EMT, hyaluronan-dependent signaling, and tumor cell plasticity. As p53 loss is prevalent in lung carcinomas, this knockout model reflects clinically relevant genetic contexts and serves as a valuable tool for evaluating CD44 as a therapeutic target or studying compensatory adaptations.
This polyclonal knockout population is suited for a breadth of functional assays. Migration and invasion can be assessed by wound healing and Transwell assays, while hyaluronan binding and flow cytometry confirm loss of CD44 surface expression. Western blotting for phospho-AKT, ERK1/2, MMP-9, and ??-catenin enables pathway analysis. Chemoresistance profiling and RNA-seq facilitate investigation of CD44-mediated drug insensitivity and global transcriptomic changes. Additional applications include cancer stem cell assays and screening of CD44-directed therapeutics. For further details or custom inquiries, please reach out to Ascent Research.
