The DNMBP Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, intended for loss-of-function studies of the DNMBP gene. This polyclonal pool contains a heterogeneous mix of cells with gene disruption at the target locus, avoiding clonal selection and capturing population-level effects. The polyclonal format is advantageous for experiments where reproducible knockout phenotypes are more important than clonal homogeneity, and it supports robust functional assays across a genetically diverse cell background.
HeLa cells, originally established from a cervical cancer biopsy, are one of the most extensively characterized human cell lines in biomedical research. They harbor integrated HPV-18 sequences, leading to sustained expression of the E6 and E7 oncoproteins, which inactivate p53 and Rb, respectively. Consequently, HeLa cells serve as a robust platform for examining oncogenic signaling and the molecular mechanisms of epithelial-derived cancers.
DNMBP (Tuba) is a scaffold protein and CDC42-specific guanine nucleotide exchange factor that coordinates actin cytoskeleton reorganization, tight junction assembly, and endocytic trafficking. At cell?Ccell contacts, DNMBP is recruited downstream of E-cadherin ligation, SRC family kinases, and PI3K signaling, where it activates CDC42 through its GEF domain. Activated CDC42 then stimulates PAK kinase and the WASP/WAVE complex to promote localized actin polymerization. DNMBP physically interacts with dynamin, the tight junction scaffold ZO-1, the polarity protein Par3, and filamentous actin, forming a nexus between adhesion, polarity, and membrane trafficking. This molecular network is essential for sustaining epithelial barrier function through the coordinated regulation of the CDC42?CRAC1?Cmyosin pathway and the proper distribution of claudins, occludin, and the E-cadherin/??-catenin adhesive complex.
Disruption of DNMBP in HeLa cells provides a model to dissect how loss of this junctional regulator affects epithelial morphology, barrier function, and invasive capacity. Although HeLa cells form incomplete tight junctions, they retain key components like ZO-1 and E-cadherin; thus, DNMBP knockout can reveal roles in residual junctional assembly, paracellular permeability, and collective cell migration. This polyclonal knockout population is suited for studying metastatic behaviors, as HeLa cells are tumorigenic and commonly used in in vitro invasion assays and xenograft models. The model also allows examination of crosstalk between adhesion signaling and actin remodeling in a transformed epithelial context.
Typical applications for these polyclonal knockout cells include immunofluorescence staining of junctional markers such as ZO-1, occludin, and E-cadherin to evaluate tight junction integrity, transwell permeability assays to quantify epithelial barrier function, and scratch wound healing assays to study collective cell migration. Co-immunoprecipitation experiments can identify DNMBP protein interaction networks, while Rho GTPase activation assays specifically measure CDC42 activity. Endocytosis assays using fluorescent ligands enable investigation of dynamin-dependent internalization pathways. These cells are also valuable for pharmacological screens aimed at identifying modulators of cell junction stability or actin cytoskeleton dynamics. For further details, customization options, and technical support, please contact Ascent Research.
