The INA Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the INA gene within the near-haploid HAP1 human cell line. This product offers a loss-of-function model for ??-internexin, a type IV intermediate filament protein essential for neuronal cytoskeletal architecture. The polyclonal format provides a heterogeneous pool of cells carrying targeted gene disruptions, enabling robust functional analyses while avoiding the biases of clonal selection. As a consistent knockout tool, it supports diverse investigations into neurofilament biology and cytoskeletal regulation.
HAP1 is a near-haploid human cell line (disomic for chromosome 8), originally derived from the chronic myeloid leukemia KBM-7 line, and is widely employed in functional genomics and knockout screens due to its haploid nature, which simplifies genetic manipulation and phenotypic interpretation. The male-derived HAP1 background offers a stable and scalable platform for CRISPR/Cas9-mediated gene editing, ensuring efficient disruption of diploid gene function. Its adaptability to high-throughput assays makes it an ideal host for modeling gene function in both cancer biology and neuronal-related processes.
The INA gene encodes ??-internexin, which co-assembles into intermediate filament networks with neurofilament light chain (NEFL), neurofilament medium chain (NEFM), neurofilament heavy chain (NEFH), and vimentin, while also interacting with nestin, peripherin, and the microtubule-associated protein MAP1B. This network is transcriptionally regulated by neurogenic factors NeuroD1 and neurogenin, and is repressed by REST/NRSF and Wnt/??-catenin signaling. INA functions downstream of these regulators to promote neurofilament assembly, axon guidance, and neuronal differentiation. Disruption of INA therefore prevents ??-internexin incorporation into filaments, impairing cytoskeletal dynamics and neurofilament organization.
In the HAP1 cellular context, INA knockout eliminates ??-internexin expression, abrogating its role in filament networks and potentially altering intracellular transport and structural integrity. Though HAP1 cells are not neuronal, they express intermediate filament system components, and this model serves as a tractable system to study cytoskeletal organization and signal transduction relevant to neurofilament dynamics. The near-haploid background enhances phenotype penetrance, allowing clear assessment via immunofluorescence for intermediate filaments, phospho-ERK/AKT analysis, and other biochemical readouts.
This INA knockout model is well-suited for investigating neuronal intermediate filament function and cytoskeletal dynamics in a simplified cellular environment. Key applications include axon regeneration studies, drug target validation for neurodegenerative disorders, and exploration of neuroblastoma and medulloblastoma biology. Researchers can employ Western blotting for INA, co-immunoprecipitation of filament complexes, neurite outgrowth assays, and RT-qPCR for neuronal differentiation markers. The polyclonal population provides a valuable resource for functional genomics screens. For additional information, please contact Ascent Research.
