The DLK1 Knockout HAP1 Polyclonal Cells are a heterogeneous population of CRISPR/Cas9-edited HAP1 cells carrying a targeted disruption of the DLK1 gene. This polyclonal knockout pool provides a versatile loss-of-function model for studying DLK1-dependent cellular processes without the need for single-cell cloning. The pooled format ensures representation of diverse editing events, enabling robust functional analyses across a genetically mixed background.
HAP1 is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line. Its near-haploid karyotype simplifies genetic manipulation, as only one allele needs to be disrupted to achieve functional knockout. HAP1 cells are also p53-deficient, which reduces stress-induced apoptosis and enhances the efficiency of genome editing. These properties make HAP1 a favored host for CRISPR-based functional genomics, high-throughput screening, and the generation of isogenic knockout collections.
DLK1 encodes a type I transmembrane protein that acts as a non-canonical ligand in the Notch signaling pathway. The protein undergoes cleavage by the metalloprotease ADAM17/TACE, releasing its extracellular domain, which competes with canonical Notch ligands (e.g., DLL1, JAG1) for binding to Notch1 and Notch2 receptors, thereby inhibiting downstream signaling. This repression leads to reduced expression of HES1 and downregulation of the adipogenic transcription factors PPAR?? and C/EBP??, ultimately blocking adipocyte differentiation. DLK1 expression is subject to paternal imprinting and is regulated by growth hormone, glucocorticoids, and the EWS/FLI1 fusion protein. In addition to ADAM17, DLK1 interacts with fibronectin and integrins, and modulates the ERK1/2 pathway, further influencing cell proliferation and survival.
In the HAP1 background, DLK1 knockout provides a clean system to dissect its role in Notch signaling and adipogenesis, unencumbered by a diploid genome. The near-haploid state and p53 deficiency facilitate the study of DLK1’s impact on cell cycle progression and apoptosis without confounding p53-dependent responses. Moreover, given the leukemic origin of HAP1, this model is particularly relevant for investigating DLK1??s contributions to cancer stem cell maintenance and tumor proliferation, as DLK1 is overexpressed in various malignancies including neuroblastoma, hepatocellular carcinoma, and small cell lung cancer.
Researchers can utilize this knockout pool for a wide range of assays, including Western blotting and RT-qPCR to confirm DLK1 disruption and assess target gene expression changes, Oil Red O staining to evaluate adipogenic differentiation capacity, and Notch reporter assays to measure pathway activity. Co-immunoprecipitation experiments can probe DLK1-Notch1 interactions, while MTS assays quantify effects on cell proliferation. Flow cytometry can be employed to monitor stem cell marker profiles. Additionally, the cells are suitable for genetic and chemical screens aimed at identifying modulators of DLK1 cleavage or Notch signaling. For technical inquiries, product customization, or ordering information, please contact Ascent Research.
