The DIAPH3 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human K-562 chronic myelogenous leukemia (CML) cell line, featuring targeted disruption of the DIAPH3 gene. This loss-of-function model enables investigation of DIAPH3-dependent actin dynamics and signaling pathways in a leukemic background while retaining genetic heterogeneity for population-level studies. The polyclonal format provides a practical tool for functional genomics without single-cell cloning.
K-562 cells were originally isolated from the pleural effusion of a 53-year-old female patient with CML in blast crisis and carry the BCR-ABL oncogenic fusion gene. These suspension-adapted cells exhibit multipotential differentiation capacity along erythroid, granulocytic, and monocytic lineages, making them a widely used model for hematopoietic differentiation and leukemia research. Their high transfectability and reproducible growth characteristics facilitate efficient CRISPR-based genome editing and subsequent phenotypic assays.
DIAPH3 encodes a member of the diaphanous-related formin family that functions as a Rho-activated actin nucleation and elongation factor. RhoA binding relieves autoinhibition, allowing DIAPH3 to promote assembly of unbranched F-actin filaments, stress fiber formation, and cell migration. DIAPH3 interacts with profilin, ??-actin, and microtubule-associated proteins APC and EB1, and its activity converges on SRF/MRTF-A-mediated transcription and focal adhesion components including vinculin and paxillin. Upstream regulators include RhoA, integrin signaling, and EGF, situating DIAPH3 at the intersection of actin cytoskeleton regulation, Rho GTPase and Hippo signaling, and adherens junction dynamics. In cochlear hair cells, DIAPH3 is essential for stereocilia maintenance, and mutations cause autosomal dominant deafness type 1 (DFNA1).
In the K-562 leukemic context, DIAPH3 disruption allows dissection of formin-mediated actin polymerization in the presence of constitutive BCR-ABL signaling, which is known to remodel the cytoskeleton and promote aberrant adhesion and migration. This knockout model is particularly suited to interrogate how DIAPH3 contributes to leukemia cell adhesion, chemotaxis, and potential egress from the bone marrow niche, processes that mimic aspects of cancer cell metastasis. By combining DIAPH3 loss with the oncogenic background of CML, researchers can examine the crosstalk between Rho GTPase-driven actin dynamics and tyrosine kinase-driven proliferative signals.
These DIAPH3 knockout K-562 polyclonal cells support a wide range of experimental applications, including Western blotting to confirm target protein depletion, phalloidin staining for F-actin visualization, flow cytometry for adhesion marker expression, and quantitative cell adhesion assays. Co-immunoprecipitation and RhoA activity assays enable investigation of DIAPH3 interactomes and upstream signaling activity. The cells are suitable for studies on actin dynamics in leukemia, the role of formins in migration and adhesion, RhoA?CDIAPH3?CSRF pathway analysis, and mechanisms of cancer metastasis. For further information, please contact Ascent Research.
