The DPYSL2 Knockout HAP1 Polyclonal Cells are a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population, designed for the targeted disruption of the human DPYSL2 gene (encoding collapsin response mediator protein 2, CRMP2) in the near-haploid HAP1 cell line. This heterogeneous pool of cells harbors diverse loss-of-function mutations, providing a robust model for functional genomics studies of CRMP2-dependent cellular processes.
HAP1 cells are a male-derived, adherent, fibroblast-like chronic myeloid leukemia (CML) cell line with a near-haploid karyotype, originally derived from the KBM-7 line. The haploid genome simplifies knockout generation, as a single allelic disruption results in a functional null phenotype, rendering HAP1 cells a preferred host for CRISPR/Cas9-mediated genetic screens and targeted gene perturbation studies.
DPYSL2 encodes CRMP2, a cytosolic phosphoprotein that promotes microtubule assembly by binding tubulin heterodimers. In response to the axonal guidance cue Semaphorin-3A (Sema3A), the Neuropilin-1/Plexin-A1 receptor complex activates a kinase cascade wherein cyclin-dependent kinase 5 (Cdk5) and glycogen synthase kinase-3?? (GSK-3??) sequentially phosphorylate CRMP2. This phosphorylation reduces CRMP2’s microtubule affinity, leading to microtubule depolymerization and growth cone collapse. Beyond microtubule regulation, CRMP2 interacts with Fyn tyrosine kinase, the endocytic adaptor Numb, kinesin-1 motor protein, and the WAVE1 actin-regulatory complex, thereby coupling extracellular signals to cytoskeletal reorganization, axonal transport, and endocytosis. Disruption of DPYSL2 therefore impacts Semaphorin signaling, axonal guidance, Rho GTPase signaling, and microtubule dynamics pathways.
In the HAP1 host context, DPYSL2 knockout offers a genetically simplified system to dissect CRMP2-dependent mechanisms without the confounding effects of a second allele. Although HAP1 cells originate from CML, they retain expression of key Semaphorin pathway components and exhibit active cytoskeletal dynamics, making them suitable for investigating fundamental processes such as neurite outgrowth, axon guidance, and cell migration. This model is particularly relevant for research into neurodegenerative disorders??including Alzheimer’s disease, schizophrenia, and amyotrophic lateral sclerosis??where CRMP2 hyperphosphorylation or misregulation is implicated, as well as for cancer studies exploring the role of CRMP2 in tumor cell invasion and metastasis.
Researchers can utilize these polyclonal knockout cells in a range of functional assays. Typical applications include neurite outgrowth immunofluorescence and growth cone collapse assays to assess axonal dynamics, microtubule co-sedimentation experiments to evaluate microtubule-binding activity, and CRMP2 phospho-specific Western blotting to monitor signaling pathway activation. Additionally, the cells are amenable to cell migration and invasion assays to investigate CRMP2??s contribution to motility in cancer, and can be deployed in high-content screening campaigns for modulators of Semaphorin signaling or Alzheimer??s drug candidates. For further information, please contact Ascent Research.
