The ANKRD44 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population engineered from the Jurkat T lymphoblastoid cell line. This product provides a loss-of-function model for functional studies of ANKRD44, the gene encoding a regulatory subunit of protein phosphatase 6 (PP6). The polyclonal knockout pool enables robust interrogation of ANKRD44-dependent signaling without clonal selection artifacts.
The Jurkat cell line is an immortalized human T lymphocyte model originally derived from an acute T cell leukemia patient. It is widely employed to investigate T cell receptor (TCR) signaling, cytokine production, HIV infection, and apoptosis regulation. Jurkat cells recapitulate key aspects of adaptive cellular immunity, including activation-induced gene expression and signaling cascades downstream of the TCR and costimulatory receptors such as CD28.
ANKRD44 functions as a scaffold that directs the PP6 catalytic subunit (PPP6C) and associated regulatory proteins PP6R1 and PP6R3 to substrate targets, most notably the kinases IKKepsilon and TBK1. By mediating dephosphorylation of these kinases, ANKRD44 modulates signaling flux through the NF-kappaB and MAPK/ERK pathways. In T cells, ANKRD44 integrates activating signals from the TCR complex, CD28 coreceptor, and inflammatory cytokines including IL-2 and TNF-alpha. Its activity influences downstream transcription factors such as NFAT, AP-1, and NF-kappaB, thereby linking phosphatase activity to gene expression programs governing immune responses.
Disruption of ANKRD44 in Jurkat T cells eliminates PP6 substrate targeting, leading to hyperphosphorylation of IKKepsilon and TBK1 and consequent dysregulation of NF-kappaB and ERK1/2 signaling. This perturbation mimics signaling abnormalities observed in T cell acute lymphoblastic leukemia, autoimmune disorders, and chronic inflammatory conditions. The knockout model thus provides a physiologically relevant context for examining how PP6-dependent dephosphorylation controls T cell activation thresholds, effector functions, and survival decisions.
Typical research applications include flow cytometric monitoring of T cell activation markers (CD69, CD25), luciferase reporter assays for NF-kappaB activity, and RT-qPCR or ELISA profiling of IL-2 and TNF-alpha production. The polyclonal ANKRD44 knockout cells are also suitable for co-immunoprecipitation experiments to identify PP6 complex components, phospho-immunoblotting to map IKKepsilon/ERK signaling dynamics, and high-throughput screening of immunomodulatory compounds or cancer immunotherapy targets. For further technical details or ordering information, please contact Ascent Research.
