The DYRK2 Knockout HAP1 Polyclonal Cells product from Ascent Research provides a CRISPR/Cas9-mediated gene-disrupted polyclonal population of HAP1 cells with targeted knockout of the DYRK2 gene. This knockout model is generated without clonal isolation, yielding a heterogeneous pool of edited cells suitable for pooled functional studies and loss-of-function experiments. The polyclonal format captures diverse editing outcomes across the population, enabling robust analysis of DYRK2-dependent phenotypes in a near-haploid genetic background.
HAP1 cells are a human near-haploid chronic myeloid leukemia (CML)-derived cell line, originally established from the KBM-7 line. These adherent, fibroblast-like cells maintain a haploid karyotype except for disomy of chromosome 8, facilitating straightforward gene disruption and functional genetic screening. The haploid nature simplifies the interpretation of knockout phenotypes by eliminating compensatory alleles, making HAP1 an ideal host for CRISPR-based gene perturbation studies. This cell line is widely used in functional genomics, drug target validation, and haploid genetic screens.
DYRK2 (dual-specificity tyrosine-phosphorylation-regulated kinase 2) is a serine/threonine kinase that functions as a tumor suppressor through multiple mechanisms. Upon DNA damage, ATM kinase activates DYRK2, which directly phosphorylates p53 at Ser46, shifting the p53 transcriptional program toward pro-apoptotic gene expression, including PUMA and BAX, thereby promoting apoptosis. DYRK2 also phosphorylates and facilitates ubiquitin-dependent degradation of oncoproteins such as c-Jun, c-Myc, and TAZ/WWTR1, thereby suppressing cell proliferation and tumor growth. The kinase interacts with ubiquitin ligases and adaptors including MDM2 and SIAH1, integrating signals from DNA damage, Hippo, and cell cycle pathways.
In the HAP1 model, disruption of DYRK2 enables investigation of its tumor-suppressive functions in a haploid setting, where loss-of-function effects are immediate and unambiguous. Researchers can interrogate DYRK2??s role in DNA damage-induced apoptosis by comparing wild-type and knockout cells exposed to genotoxic agents, with readouts such as p53 phosphorylation at Ser46 and caspase-3 cleavage. The knockout pool can also be used to study DYRK2-mediated regulation of protein stability for c-Myc and TAZ, as well as downstream transcriptional effects on proliferation and survival. The haploid background minimizes genetic redundancy, allowing for clear dissection of DYRK2 signaling networks.
This polyclonal DYRK2 knockout cell product is valuable for diverse experimental applications in cancer biology, DNA damage response, Hippo signaling, and drug sensitivity testing. Typical assays include Western blotting for p-p53(Ser46), p53, and cleaved caspase-3; apoptosis assays; DNA damage foci analysis; cell proliferation measurements; co-immunoprecipitation to assess DYRK2-p53 interactions; RT-qPCR for PUMA, BAX, and p21; and drug treatments with doxorubicin or etoposide combined with proteasome inhibitor MG132 or cycloheximide chase for ubiquitination studies. This model supports mechanistic dissection of tumor suppression and evaluation of therapeutic vulnerabilities. For additional information, please contact Ascent Research.
