The CASKIN2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout population targeting the CASKIN2 gene in the near-haploid HAP1 human cell line. This product comprises a heterogeneous pool of cells with diverse loss-of-function mutations, enabling robust functional studies without clonal bias. The polyclonal format is particularly suited for pooled phenotypic assays and protein interaction analyses requiring population-level gene disruption.
Derived from the chronic myeloid leukemia line KBM-7, HAP1 cells are a near-haploid fibroblast-like line widely used for genetic knockout studies. Their single-allele genome simplifies CRISPR editing and phenotypic interpretation, as disruption of one copy abolishes gene expression. HAP1 cells support standard transfection and culture protocols, providing a versatile platform for investigating signaling, adhesion, and cytoskeletal mechanisms.
CASKIN2 encodes a synaptic scaffolding protein that bridges the adhesion molecule CASK to the actin cytoskeleton via direct interaction with the liprin-?? family member PPFIA1. The CASK-CASKIN2-liprin complex integrates synaptic adhesion with actin filament dynamics, thereby influencing dendritic spine morphology and synaptic plasticity. Additionally, CASKIN2 associates with SHANK scaffold proteins, positioning it within the postsynaptic density network. Through these interactions, CASKIN2 regulates actin reorganization downstream of synaptic adhesion signals, linking membrane-associated guanylate kinases to cytoskeletal remodeling essential for proper synapse formation and maintenance.
Despite their non-neuronal origin, HAP1 cells provide an effective model for dissecting the core biochemical functions of CASKIN2. The near-haploid background ensures complete loss of protein expression across the knockout population, facilitating clear readouts in protein interaction and cytoskeletal assays. This system enables detailed mapping of CASKIN2-dependent complexes, including interactions with CASK, PPFIA1, and SHANK proteins, without the complexity of neuronal differentiation. Thus, it serves as a valuable tool for studying actin dynamics and scaffold protein assembly relevant to synaptic architecture.
Researchers can employ western blotting and co-immunoprecipitation to validate CASKIN2 knockout and probe altered protein interactions within the CASK-liprin-actin axis. Immunofluorescence microscopy and actin polymerization assays provide direct visualization and quantification of cytoskeletal changes. Cell adhesion assays further reveal functional consequences on substrate attachment. This knockout population supports functional genomics screens, proteomics, and studies of synaptic scaffolding mechanisms relevant to neurodevelopmental disorders. For additional information, please contact Ascent Research.
