The CD44 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the K-562 cell line, engineered to disrupt the CD44 gene. This product offers a heterogeneous pool of cells with targeted loss-of-function mutations, providing a robust model for studying CD44-dependent biological processes without the clonal biases inherent in single-cell-derived lines. The polyclonal format preserves the genetic diversity of the edited population, enabling the investigation of CD44 function in a more physiologically relevant context.
The K-562 host cell line was established from a 53-year-old female with chronic myelogenous leukemia (CML) in blast crisis. K-562 cells are widely employed as a model system for hematopoietic differentiation and leukemia, particularly for studying CML biology and the BCR-ABL oncogenic signaling. These cells exhibit characteristics of undifferentiated blast cells and are extensively used to investigate mechanisms of leukemogenesis, drug resistance, and signal transduction in a myeloid lineage background.
CD44 encodes a transmembrane glycoprotein that functions as the primary receptor for hyaluronic acid, but also interacts with osteopontin, collagen, and MMP-9. Upon ligand engagement, CD44 recruits ERM proteins (ezrin, radixin, moesin) to link the receptor to the actin cytoskeleton and initiates downstream signaling cascades. Specifically, CD44 activates the PI3K/Akt and MAPK/ERK pathways, and stimulates small GTPases including Rac1 and Cdc42, leading to cytoskeletal reorganization and transcriptional regulation. CD44 expression is induced by cytokines such as TNF-?? and IL-1?? through transcription factors NF-??B, AP-1, and Egr-1, and is post-transcriptionally regulated by miR-34a. Downstream, CD44 modulates the expression of MMP-9 and ??-catenin, and influences processes such as cell adhesion, migration, and proliferation.
In the context of K-562 chronic myelogenous leukemia cells, CD44 is known to mediate critical interactions with the bone marrow microenvironment through hyaluronic acid binding, contributing to leukemic cell adhesion, homing, and survival. Disruption of CD44 in this model allows for the dissection of its role in CML pathogenesis, including its potential involvement in drug resistance mechanisms and maintenance of leukemic stem cell properties. Given that K-562 cells represent blast crisis phase CML, this knockout model is particularly valuable for studying the aggressive stage of the disease and evaluating CD44 as a therapeutic target.
This CD44 knockout cell population is suitable for a range of research applications. Investigators can use flow cytometry to verify CD44 surface protein loss and adhesion assays on hyaluronic acid-coated surfaces to assess CD44-mediated attachment. Migration and invasion assays can elucidate the role of CD44 in leukemic cell motility, while Western blotting for phospho-Akt and phospho-ERK reveals CD44-mediated pathway activation. RT-qPCR can quantify transcriptional changes in downstream targets such as MMP-9. Additionally, proliferation and drug sensitivity assays can evaluate the impact of CD44 loss on cell growth and response to tyrosine kinase inhibitors. For further details, please contact Ascent Research.
