The DMPK Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population engineered for targeted disruption of the DMPK gene in the human near-haploid HAP1 cell line. This polyclonal pool encompasses a heterogeneous mixture of edited alleles, providing a robust loss-of-function model for functional genomics studies. The product is supplied as a live cell population and is optimized for downstream applications ranging from biochemical assays to high-throughput phenotypic screening. By abolishing DMPK expression across a diversified genetic background, the cells enable dissection of DMPK-dependent signaling without the confounding effects of clonal variation, making them suitable for investigating gene function in a physiologically relevant cellular context.
The HAP1 host cell line is a near-haploid human cell model derived from the KBM-7 chronic myelogenous leukemia line, originally isolated from a male patient. These adherent cells retain a predominantly haploid karyotype, which facilitates straightforward genetic manipulation and knockout screening by minimizing functional redundancy from diploid alleles. HAP1 cells are widely employed in functional genomics, drug target validation, and CRISPR-based arrayed screens due to their stable growth characteristics and the ease of establishing homozygous disruptions. Their leukemic origin also provides a unique background for studying signaling pathways intersecting with hematopoietic malignancies and cancer cell biology.
DMPK encodes a serine/threonine kinase that critically regulates actin-myosin contractility and alternative pre-mRNA splicing. The kinase functions downstream of upstream regulators including MyoD, MEF2, Ca2?/calmodulin, RhoA, and integrin signaling, and directly phosphorylates downstream targets such as MYPT1 (PPP1R12A) and CELF1 (CUG-BP1). Phosphorylation of MYPT1 modulates myosin phosphatase activity, thereby controlling myosin light chain (MLC) phosphorylation and actomyosin dynamics. DMPK-mediated phosphorylation of CELF1 influences its RNA-binding activity, impacting alternative splicing of targets like TNNT2 and CLCN1. DMPK also interacts with MBNL1, HSP90, HSF1, and MYPT1, forming regulatory complexes that coordinate splicing and cytoskeletal remodeling. These molecular connections place DMPK at the nexus of actin cytoskeleton regulation, calcium signaling, muscle contraction, insulin signaling, and RNA processing pathways.
In HAP1 cells, DMPK knockout disrupts the kinase-dependent phosphorylation cascade, impairing myosin phosphatase regulation and CELF1-driven splicing programs. This perturbation recapitulates key molecular hallmarks of myotonic dystrophy type 1, including aberrant alternative splicing and compromised muscle contractility. The near-haploid background enhances the ability to link genotype to phenotype, offering a simplified system for studying DMPK-related pathologies such as cardiac arrhythmia, myotonia, and muscle atrophy. As HAP1 cells lack the full complement of muscle-specific factors, they provide a reductionist platform for dissecting core DMPK mechanisms without the complexity of differentiated muscle environments, while still supporting relevant actin-myosin signaling events.
This knockout model is ideally suited for myotonic dystrophy type 1 modeling, muscle contraction studies, and alternative splicing analysis. Researchers can employ RNA-seq and splicing reporter assays to assess splicing changes in targets like TNNT2 and CLCN1, or use phospho-MLC assays and cell migration assays to evaluate actomyosin dynamics. High-content imaging and immunofluorescence enable visualization of cytoskeletal alterations, while co-immunoprecipitation and western blotting facilitate study of DMPK interactors such as MBNL1 and MYPT1. The polyclonal population is also amenable to high-throughput drug screening, enabling identification of small molecules that modulate DMPK-related pathways. For further information, please contact Ascent Research.
