The INA Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T-lymphocyte line, engineered for targeted disruption of the INA gene encoding alpha-internexin. This loss-of-function model enables researchers to investigate INA biology in a non-neuronal context or to serve as a negative control in experiments where neuronal intermediate filament expression is undesirable. The polyclonal nature of the edited pool preserves genetic diversity, offering a robust tool for high-throughput screens and pooled functional genomics assays.
The Jurkat cell line is an immortalized human T-lymphocyte line originating from a patient with acute T-cell leukemia, widely employed for studies of adaptive immunity, T-cell receptor signaling, and cytokine production. Its well-characterized signaling networks and favorable growth characteristics make it a reliable host for CRISPR/Cas9-mediated genome engineering, enabling precise gene disruption studies in a hematopoietic context.
Alpha-internexin, encoded by INA, is a type IV neuronal intermediate filament protein that co-assembles with neurofilament light (NEFL), medium (NEFM), and heavy (NEFH) subunits to form the axonal cytoskeleton, essential for axon radial growth and structural integrity. Transcription factors Neurogenin, NeuroD, and Brn3, along with NGF-MAPK signaling, promote INA expression during neuronal differentiation. Alpha-internexin interacts with microtubules and is phosphorylated by Cdk5 and GSK-3??, regulating filament assembly and axonal transport. Downstream, INA contributes to neurofilament network formation and mechanical stabilization of neurons, operating within a pathway that includes MAPT (tau) and tubulin.
In Jurkat cells, which lack endogenous INA expression, this knockout serves primarily as a negative control for neuronal gene studies or for reconstitution experiments probing intermediate filament assembly. The polyclonal knockout pool mimics cellular heterogeneity, facilitating population-level analyses. Additionally, any non-canonical roles of neurofilament proteins in immune cells can be explored using this model, potentially revealing novel aspects of cytoskeletal biology in T lymphocytes.
This product supports diverse applications, including use as a negative control in neuronal-like differentiation assays, co-immunoprecipitation for validating alpha-internexin interactions with NEFL, NEFM, NEFH, or microtubules, and genomic DNA sequencing or RT-qPCR to confirm gene disruption. Immunofluorescence staining and Western blotting can assess protein expression and localization in knockout versus wild-type cells. The cells are suitable for protein interaction studies, high-content screening for neurofilament modulators, and neurodegenerative disease research focusing on amyotrophic lateral sclerosis and Charcot-Marie-Tooth disease. For further technical details, please contact Ascent Research.
