The DLGAP4 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human K-562 chronic myelogenous leukemia cell line. These cells harbor targeted disruption of the DLGAP4 gene, resulting in loss of functional DLGAP4 protein. The polyclonal format provides a heterogeneous knockout pool that minimizes clonal selection biases and captures the average effect of DLGAP4 ablation. This product is designed for advanced research into scaffold protein functions in cancer cell signaling, adhesion, and proliferation.
The host K-562 cell line was established from pleural effusion of a 53-year-old female with CML in blast crisis and is characterized by the BCR-ABL1 fusion oncogene. K-562 cells exhibit autonomous growth, survival signaling, and serve as a canonical model for studying leukemia biology, tyrosine kinase inhibitor responses, and downstream pathways such as AKT/mTOR. Their suspension culture and well-defined genetic background facilitate high-throughput genetic and pharmacological screens.
DLGAP4 (Discs Large-Associated Protein 4) belongs to the SAPAP family of scaffold proteins that couple DLG family MAGUKs to signaling effectors and the cytoskeleton. DLGAP4 directly interacts with DLG4 (PSD-95), DLG1, and the SHANK1/2 scaffolding proteins to assemble macromolecular complexes at the plasma membrane. Through these interactions, DLGAP4 recruits and facilitates phosphorylation of AKT, leading to mTOR activation and promotion of cell proliferation and survival. The DLGAP4 gene is transcriptionally regulated by the E2F1 transcription factor. Disruption of DLGAP4 in K-562 cells is predicted to destabilize DLG4-anchored scaffolds, thereby impairing AKT/mTOR signal transduction, reducing adhesion, and attenuating proliferative capacity.
In the BCR-ABL1-driven K-562 leukemia background, DLGAP4 knockout provides a unique system to dissect the contribution of scaffold-organized signaling to oncogenic phenotypes. K-562 cells rely on constitutive activation of AKT/mTOR and adhesion pathways for survival and clonal expansion. Loss of DLGAP4 may compromise membrane-proximal signaling complexes that integrate BCR-ABL1 signals with downstream effectors, potentially revealing synthetic vulnerabilities. This polyclonal model permits assessment of DLGAP4 roles in cell migration, apoptosis, and proliferation without artifacts from single-clone selection, making it suitable for functional genomics and drug response studies.
Typical research applications include elucidating DLGAP4 function in AKT/mTOR signaling via Western blot analysis of phospho-AKT and downstream targets, and characterizing protein interaction networks using co-immunoprecipitation of DLG4 and associated partners. Leukemia-focused studies can employ proliferation assays (MTS/BrdU), flow cytometry for apoptosis (Annexin V/PI), and Transwell migration assays. RNA-seq transcriptomics can uncover global gene expression changes upon DLGAP4 loss. This polyclonal knockout population is also valuable for validating DLGAP4 as a therapeutic target and for identifying scaffold-dependent signaling nodes in cancer. For additional product details or experimental support, please contact Ascent Research.
