The DYRK2 Knockout HEK293T Polyclonal Cells product provides a heterogeneous population of HEK293T cells harboring CRISPR/Cas9-mediated disruption of the DYRK2 gene. This polyclonal knockout model disrupts endogenous DYRK2 expression, generating a loss-of-function background suitable for investigating the kinase??s roles in apoptosis, cell cycle progression, and protein degradation. The cells are supplied as a viable polyclonal pool, preserving genetic diversity while eliminating DYRK2 function, making them a flexible tool for pathway analysis, functional genomics, and phenotypic screening in a widely used human cell background.
HEK293T cells are a derivative of the HEK293 human embryonic kidney line that stably expresses the SV40 large T antigen. This enables episomal replication of plasmids containing the SV40 origin of replication, facilitating high-level protein expression and efficient viral production. The cells exhibit adherent epithelial morphology and are extensively employed in signal transduction studies, recombinant protein production, and lentivirus packaging. Their robust growth and ease of transfection make them an ideal host for CRISPR-based gene editing, and the DYRK2 knockout polyclonal population retains these advantages while specifically ablating a key regulator of stress-induced apoptosis.
DYRK2 encodes a serine/threonine kinase with crucial functions in the DNA damage response. Activation occurs downstream of ATM and ATR kinases, often via CHK2, leading to phosphorylation of p53 at Ser46. This modification shifts p53 toward transactivation of pro-apoptotic genes such as PUMA and BAX, thereby promoting mitochondrial apoptosis. DYRK2 also phosphorylates c-Jun and c-Myc, targeting them for ubiquitin-mediated degradation through interactions with the EDD E3 ligase and DDB1-CUL4 complex, which suppresses cell proliferation. Additionally, DYRK2 modulates Hippo signaling by phosphorylating TAZ (WWTR1), and influences NF-??B pathway activity, positioning it at the intersection of multiple tumor-suppressive networks.
In the HEK293T background, DYRK2 knockout disrupts p53-mediated apoptotic signaling and alters cell cycle control. Although the SV40 large T antigen inactivates p53 by direct binding, residual p53 functions and p53-independent actions of DYRK2 remain relevant. The polyclonal nature of this cell population offers a physiologically diverse loss-of-function model, enabling researchers to study overall gene requirement without clonal artifacts. It is particularly suited for examining how DYRK2 loss affects DNA damage-induced cell death, ubiquitin-mediated proteolysis, and cross-talk with the Hippo pathway, providing a valuable system for mechanistic studies in a cell line that otherwise supports robust molecular biology workflows.
Key applications include apoptosis research using flow cytometry with Annexin V/PI staining, cell cycle analysis, and DNA damage response assays following genotoxic stress. Western blotting and RT-qPCR can assess changes in DYRK2 downstream targets such as phospho-p53 (Ser46), c-Jun, and c-Myc. Co-immunoprecipitation experiments enable interrogation of DYRK2-containing complexes, while luciferase reporter assays for p53 activity quantify transcriptional outcomes. The cells are also suitable for drug screening campaigns targeting the DYRK2-p53 axis in colorectal, breast, and lung cancers, as well as neurodegenerative disorder studies. For further technical details, please contact Ascent Research.
