The ANK3 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the human HAP1 cell line, carrying a targeted disruption of the ANK3 gene. Unlike clonal knockout lines, this polyclonal format preserves the genetic diversity inherent to the edited pool, enabling researchers to study ANK3 loss-of-function in a representation of heterogeneous editing outcomes. This model is well-suited for initial phenotypic screening, pooled functional analysis, and studies where clonal effects may mask subtle phenotypes.
HAP1 cells are a near-haploid human chronic myeloid leukemia cell line established from the KBM-7 line, exhibiting fibroblast-like adherent growth. Their near-haploid genome simplifies CRISPR/Cas9-based gene targeting and reduces functional redundancy, making them an optimal host for genetic screens and the generation of knockout models. This background is particularly advantageous for studying genes involved in cytoskeletal and adhesion functions, as it provides a clean genetic background for assessing morphological and migratory changes.
The ANK3 gene encodes ankyrin-G, a multidomain scaffold protein that organizes ion channels and cell adhesion molecules at specific membrane domains. Ankyrin-G directly interacts with voltage-gated sodium channels (SCN1A/Nav1.2, SCN8A/Nav1.6), potassium channels (KCNQ2, KCNQ3), and cell adhesion proteins such as E-cadherin and L1CAM, linking them to the spectrin-actin cytoskeleton via ??II-spectrin (SPTBN2) and actin (ACTB). This scaffold is critical for the clustering and retention of these molecules at the axon initial segment, nodes of Ranvier, and intercalated discs. Ankyrin-G expression is regulated by transcription factors REST and SP1, as well as by WNT signaling and neuronal activity, positioning it as a downstream effector of multiple signaling cascades. Additionally, ankyrin-G interacts with dynactin to couple the ankyrin network to dynein-mediated axonal transport.
In the HAP1 cellular context, ANK3 knockout disrupts ankyrin-G-mediated cytoskeletal and adhesion networks, providing a platform to dissect these processes independently of neuronal specialization. HAP1 cells retain key components of adherens junctions and the cortical actin cytoskeleton, allowing assessment of how loss of ankyrin-G affects cell spreading, migration, and junctional stability. This model thus enables the study of ankyrin-G??s role in maintaining membrane protein localization and cytoskeletal architecture, with potential implications for understanding diseases such as Brugada syndrome, where ankyrin-G dysfunction impairs cardiac intercalated disc organization, and for exploring its contributions to cancer cell motility.
Researchers can utilize these polyclonal ANK3 knockout HAP1 cells in diverse experimental workflows including functional genomics, drug target validation, and mechanistic studies of ANK3-related psychiatric and cardiac disorders. Suited assays include western blotting to confirm loss of ankyrin-G protein, immunofluorescence microscopy to visualize disrupted ??II-spectrin and E-cadherin localization, and migration assays to quantify changes in cell motility. Co-immunoprecipitation experiments can further explore the altered interactome of residual ankyrin-G or its binding partners. For technical inquiries or to discuss custom knockout services, please contact Ascent Research.
