The DPYSL5 Knockout HT29 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DPYSL5 gene. This loss-of-function model allows investigation of DPYSL5 (CRMP5) function in the well-characterized HT29 human colorectal adenocarcinoma epithelial cell line. Suitable for studies without single-cell cloning, the polyclonal population maintains the inherent genetic diversity of the parental line.
The HT29 cell line, derived from a primary colon adenocarcinoma of a 44-year-old female, exhibits epithelial morphology and serves as a widely used model for intestinal epithelial biology and colorectal cancer research. Its robust signaling networks and well-documented behavior make it an ideal host for studying cancer cell migration and invasion.
DPYSL5 encodes collapsin response mediator protein 5 (CRMP5), a critical mediator of semaphorin signaling that orchestrates cytoskeletal dynamics. CRMP5 functions downstream of the Sema3A/neuropilin-1/plexin-A receptor complex and is regulated by upstream kinases including GSK3beta, as well as reelin signaling. It directly interacts with tubulin and actin, and associates with CDK5 and other CRMP family members to modulate microtubule assembly, actin filament reorganization, and focal adhesion turnover. Through its engagement of Rho GTPase pathways, specifically RhoA and ROCK, DPYSL5 translates extracellular guidance cues into changes in cell shape and motility. In colorectal cancer, DPYSL5 has been implicated in enhancing cell migration and invasion, highlighting its potential role in metastatic progression.
In the HT29 colorectal adenocarcinoma background, knockout of DPYSL5 is expected to disrupt semaphorin-induced cytoskeletal remodeling, leading to impaired cell migration and invasion. This model offers a relevant system to investigate how loss of DPYSL5 affects colorectal cancer cell behavior, particularly in the context of epithelial-to-mesenchymal transition and metastatic spread. The polyclonal knockout population preserves the genetic heterogeneity of the parental HT29 line, which may more accurately reflect the diversity of tumor cell responses observed in vivo. Thus, it provides a valuable tool for studying the molecular mechanisms linking semaphorin signaling to actin and microtubule dynamics in colorectal cancer.
This knockout model is well-suited for functional studies using scratch wound healing assays to assess collective cell migration, transwell migration/invasion assays to measure chemotactic and invasive potential, and immunofluorescence microscopy to visualize changes in the actin and tubulin cytoskeleton. Western blotting for total and phosphorylated CRMP5 can be used to confirm pathway disruption, while RT-qPCR provides validation of knockout efficiency. Key applications include dissecting semaphorin-plexin signaling mechanisms in colorectal cancer, conducting functional analysis of CRMP family proteins in epithelial cells, and screening for compounds that inhibit metastatic behavior. For detailed product information or to inquire about custom modifications, please reach out to Ascent Research.
