The DPYSL3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population produced by disrupting the DPYSL3 gene in HEK293T host cells. This heterogeneous pool of knockout cells provides a loss-of-function model to study the roles of DPYSL3 (also known as collapsin response mediator protein 4, CRMP4) in semaphorin signaling, cytoskeletal regulation, and related cellular processes. The polyclonal format captures a spectrum of gene-edited alleles, avoiding clonal selection artifacts, and represents a versatile tool for pathway analysis and functional screening.
The host cell line, HEK293T, is an SV40 large T-antigen-transformed human embryonic kidney epithelial cell line. It grows adherently and is widely adopted for its high transfection efficiency and robust protein expression capabilities. HEK293T cells are a standard platform for recombinant protein production, lentiviral packaging, and transient expression assays. Their amenability to genetic manipulation and established use in signal transduction studies make them a suitable background for dissecting the DPYSL3 signaling axis.
DPYSL3/CRMP4 is a phosphoprotein that mediates semaphorin-induced axon guidance and cytoskeletal reorganization. Upon activation by Semaphorin 3A (Sema3A) through the Neuropilin-1/Plexin-A receptor complex, DPYSL3 is regulated by upstream kinases including GSK-3?? and Cdk5. It interacts with cytoskeletal components such as tubulin and actin, as well as family members CRMP1 and CRMP2, the kinase GSK-3??, and the tyrosine kinase Fyn. Downstream, DPYSL3 modulates microtubule polymerization, actin dynamics, cell adhesion, and migration. It functions within a network that includes Rho GTPases to control growth cone collapse and neuronal polarity.
In the HEK293T context, loss of DPYSL3 disrupts the semaphorin signaling cascade, providing a clean background to study ligand-dependent cytoskeletal changes. Although HEK293T cells are of non-neuronal origin, they express many core signaling components and are permissive for ectopic expression of neuronal receptors, enabling reconstitution of axon guidance pathways. This knockout pool is particularly relevant for investigating cancer cell migration, as DPYSL3 has been implicated in tumor invasion and metastasis. The polyclonal nature allows researchers to assess population-level responses while circumventing clonal biases.
This knockout model supports a wide range of experimental approaches. Western blotting and RT-qPCR confirm the absence of DPYSL3 protein and mRNA, respectively. Immunofluorescence can visualize altered cytoskeletal organization, while cell migration and invasion assays quantitatively assess functional consequences. Phospho-signaling analysis following Sema3A stimulation reveals pathway activation defects. Typical applications include screening for modifiers of semaphorin signaling, investigating CRMP4-dependent tumor cell motility, and modeling neurodevelopmental disturbances. For further details or to inquire about this product, please contact Ascent Research.
