The ANKRD28 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population derived from the Jurkat T lymphocyte leukemia line, engineered to disrupt the ANKRD28 gene. This heterogeneous knockout pool enables loss-of-function studies of ANKRD28, an inhibitory subunit of the protein phosphatase 6 (PP6) holoenzyme, within a well-characterized model of T cell receptor (TCR) signaling and apoptosis. The product provides a powerful tool for functional genomics and mechanistic investigations into PP6-dependent regulatory networks.
The parental Jurkat cell line is an immortalized human T lymphocyte established from a male acute T cell leukemia patient in 1976. This line has been extensively employed to dissect TCR-mediated signaling cascades, activation-induced cell death, and associated checkpoints. Its robust proliferation and well-mapped signal transduction pathways make Jurkat cells an optimal background for assessing oncogenic and immune signaling perturbations resulting from targeted gene knockouts.
ANKRD28 serves as a critical regulatory subunit that binds the catalytic subunit PPP6C, alongside scaffold proteins PPP6R2 and PPP6R3, to suppress PP6 phosphatase activity. Disruption of ANKRD28 abrogates this inhibition, leading to constitutive PP6 activation and enhanced dephosphorylation of downstream substrates including IRF3, CHK1, and DNA-PKcs. Consequently, signaling through the NF-??B (p65) transcription factor and the MAPK cascades (ERK/MAPK1, JNK/MAPK8, p38/MAPK14) is modulated. Upstream inputs from TNF?? signaling, DNA damage signals, and TCR activation converge on ANKRD28, positioning it as a node that coordinates inflammation, cell cycle progression, and DNA repair responses.
In the Jurkat context, ANKRD28 knockout creates a relevant system for probing the intersection of TCR?CZAP70?CNF-??B signaling and PP6-mediated regulation. Loss of ANKRD28 function may attenuate NF-??B?Cdependent transcriptional programs, alter MAPK-driven proliferation or apoptosis, and compromise DNA damage checkpoints, reflecting the gene’s link to melanoma, autoimmune disorders, and inflammatory diseases. This model thus enables dissection of how PP6 deregulation influences T lymphocyte fate decisions and oncogenic processes.
Applications of these polyclonal knockout cells span T cell signaling analysis, DNA damage response research, drug target validation, and autoimmune disease modeling. Representative experimental approaches include western blotting for phospho-IRF3 and phospho-CHK1, flow cytometry for activation markers CD69 and CD25, NF-??B luciferase reporter assays, co-immunoprecipitation of PP6 complex components, ??H2AX immunofluorescence, cell cycle analysis, and Annexin V/PI apoptosis assays. Transcriptomic profiling by RNA-seq can further uncover global expression changes. For further information, please contact Ascent Research.
