The DMPK Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney cell line. This product features targeted disruption of the DMPK gene, which encodes a serine/threonine protein kinase, creating a heterogeneous pool of edited cells suitable for loss-of-function studies. The polyclonal format provides a robust model for investigating DMPK-dependent signaling without clonal isolation, capturing the variability inherent in CRISPR-mediated gene disruption. Users can perform transient or stable assays requiring DMPK deficiency, such as kinase activity profiling, protein interaction mapping, and functional complementation.
The parental HEK293T cell line is an immortalized human embryonic kidney epithelial derivative of HEK293, stably expressing SV40 large T antigen, which enables episomal replication of plasmids containing the SV40 origin, leading to high transfection efficiency and robust recombinant protein expression. HEK293T cells are amenable to diverse genetic manipulations and biochemical assays, making them optimal for generating knockout models. Their epithelial origin and flat morphology facilitate imaging-based analyses, while rapid doubling time supports high-throughput screening.
DMPK is a serine/threonine kinase that regulates actomyosin contractility by phosphorylating myosin regulatory light chain (MYL9) and myosin phosphatase target subunit 1 (MYPT1). It also modulates ion channel activity via phospholemman (PLN) phosphorylation, influencing calcium handling. Upstream, MEF2, SRF, and ROCK activate DMPK transcriptionally and post-translationally. DMPK interacts with HSPB1, HSPA8, actin, ??-actinin, and calmodulin, integrating cytoskeletal dynamics and stress responses. This kinase contributes to Wnt/??-catenin, MAPK/ERK, and insulin signaling pathways.
In HEK293T cells, DMPK disruption offers a simplified system to study kinase-dependent functions without tissue-specific confounders. The robust proliferation of HEK293T enables efficient expansion of the polyclonal pool for large-scale studies. Despite being non-muscle, these cells express actomyosin and ion channel components, allowing investigation of DMPK’s conserved roles in contractility and ion homeostasis. This model is valuable for exploring DMPK scaffolding functions and signaling networks linked to myotonic dystrophy type 1, cardiac defects, and cataracts.
These polyclonal knockout cells are ideal for phosphoproteomic profiling to identify novel DMPK substrates, screening small-molecule modulators, and co-immunoprecipitation studies to map protein interaction networks. Researchers can confirm DMPK knockout by Western blotting and assess kinase activity using phospho-specific antibodies against known targets such as MYPT1 and PLN. Actin cytoskeleton staining reveals contractility defects, and calcium imaging or proliferation assays evaluate ion handling and growth control. Functional complementation by re-expression of wild-type or mutant DMPK enables structure-function analysis. For further information, please contact Ascent Research.
