The HTT Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte line. This product features disruption of the HTT gene, eliminating expression of the huntingtin scaffold protein. As a polyclonal pool, these cells harbor a heterogeneous mix of gene-edited alleles, providing a robust loss-of-function model suitable for high-throughput studies. The cells enable investigation of huntingtin biology in an immune cell context and serve as a versatile platform for biochemical and pharmacological assays.
Jurkat is a human T cell leukemia cell line originally isolated from the peripheral blood of a 14-year-old male with acute T cell leukemia. Widely employed as a model for T cell receptor signaling, apoptosis, and leukemogenesis, Jurkat cells grow rapidly and tolerate genetic manipulation well. They express key TCR signaling components such as Lck, ZAP-70, and NFAT, and respond to phorbol esters and calcium ionophores. These features make Jurkat an excellent host for CRISPR/Cas9-based knockout models to dissect immune signaling and cancer biology.
Huntingtin (HTT) is a large scaffold protein that coordinates intracellular trafficking, transcriptional regulation, and anti-apoptotic signaling. It interacts with HAP1, HIP1, clathrin, and dynein to regulate vesicle movement, and is phosphorylated by AKT1 and CDK5. HTT modulates gene expression by interacting with the REST/NRSF complex, thereby controlling BDNF transcription, and also influences Wnt/??-catenin and Notch signaling. Additionally, HTT binds LC3 and p62/SQSTM1 to facilitate autophagy and suppresses caspase-3/7-mediated apoptosis. Knockout of HTT disrupts these networks, impairing trafficking, autophagy, and cell survival pathways.
In Jurkat T cells, HTT knockout creates a non-neuronal model for investigating huntingtin function. T lymphocytes depend on autophagy for homeostasis and activation, and on caspase-mediated apoptosis for immune regulation, making them susceptible to HTT loss. Disrupted intracellular trafficking can alter T cell receptor internalization and downstream signaling. Moreover, aberrant REST/NRSF activity may cause misexpression of neuronal genes in these lymphoid cells. This system allows researchers to dissect cell-autonomous, huntingtin-dependent processes in a dividing, immune-competent background, complementing studies in neurons.
These HTT knockout polyclonal cells are suited for high-throughput drug screening to identify modulators of huntingtin-related pathways, functional proteomics to map HTT interaction partners, and transcriptomic analyses such as RNA-seq. Typical assays include Western blotting for HTT protein verification, RT-qPCR for mRNA levels, flow cytometry for apoptosis (Annexin V/PI and caspase activation), immunofluorescence for LC3 puncta, and autophagy flux measurement via p62 degradation. For further technical information, please contact Ascent Research.
