The DPYSL5 Knockout HAP1 Polyclonal Cells are a polyclonal population of HAP1 cells engineered to carry a CRISPR/Cas9-mediated disruption of the DPYSL5 gene. This product provides a pooled loss-of-function model, eliminating the need for single-cell cloning and enabling interrogation of DPYSL5-dependent phenotypes across a population of edited cells.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype ensures that most genes, including DPYSL5 on chromosome 22, exist in a single copy, thereby simplifying knockout studies by avoiding the compensation often observed in diploid cells. HAP1 cells maintain robust growth in culture and are widely used for functional genomics, drug screening, and high-content imaging.
DPYSL5 encodes CRMP5, a member of the collapsin response mediator protein family that serves as a downstream effector in semaphorin-plexin signaling. Upon Semaphorin3A stimulation, the Neuropilin-1/Plexin-A receptor complex activates Fyn kinase, leading to Cdk5- and GSK3??-mediated phosphorylation of CRMP5. Phosphorylated CRMP5 interacts with tubulin and actin, promoting microtubule reorganization and growth cone collapse. CRMP5 also forms complexes with other CRMP family members (CRMP1, CRMP2, CRMP4) and with cofilin, linking it to RhoA and Rac1 GTPase signaling pathways.
In the HAP1 context, disruption of DPYSL5 offers a clean genetic background to dissect CRMP5-mediated cytoskeletal dynamics without the confounding influence of neuronal-specific factors. The knockout pool can be used in reconstitution assays to map phosphorylation-dependent interactions, or to screen for chemical modulators of the semaphorin pathway. Additionally, because HAP1 cells exhibit migratory behavior, the knockout model is suitable for studying the role of CRMP5 in cancer cell migration and invasion.
Typical applications include immunofluorescence staining to assess microtubule and actin organization, quantitative Western blotting of phospho-CRMP5 and downstream targets, and functional assays such as neurite outgrowth following neuronal differentiation or wound-healing migration assays. The polyclonal format ensures robust, reproducible phenotypes while reflecting the heterogeneity inherent in CRISPR-edited populations. For further information or to request a technical consultation, please contact Ascent Research.
