The DPYSL3 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the DPYSL3 gene in the HCT 116 human colorectal carcinoma cell line. This loss-of-function model enables investigation of dihydropyrimidinase-like 3 (CRMP4) in cancer biology and signal transduction. As a polyclonal pool, it captures diverse editing events, avoiding clonal selection biases.
HCT 116 is a colorectal adenocarcinoma epithelial line harboring KRAS (G13D) and PIK3CA (H1047R) mutations, which constitutively activate MAPK and PI3K/AKT pathways. This widely used model offers a tumorigenic background for studying oncogenic signaling and therapeutic responses.
DPYSL3 encodes CRMP4, a collapsin response mediator protein that integrates semaphorin and reelin signals to regulate cytoskeletal dynamics. CRMP4 acts downstream of SEMA3A?CNRP1/PLXNA1 receptor complexes and is phosphorylated by GSK3?? and Fyn. It directly binds tubulin and actin and forms complexes with CRMP1 and CRMP2, modulating microtubule polymerization and actin filament remodeling. Through Rho GTPase signaling, CRMP4 controls RhoA, cofilin, and myosin light chain 2 (MLC2) activity, thereby governing cell migration, adhesion, and morphological plasticity.
In colorectal cancer, DPYSL3 loss is linked to heightened invasive potential. In HCT 116 cells with activating KRAS and PIK3CA mutations, CRMP4 depletion may uncover dependencies on RhoA-driven actomyosin contractility and cofilin-mediated actin severing. This model thus facilitates dissection of how DPYSL3 loss cooperates with established driver mutations to enhance migration, confer anoikis resistance, and modulate drug sensitivity.
Key applications include quantitative wound healing and Transwell invasion assays to measure migration and invasion, complemented by immunofluorescence staining of F-actin and microtubules for cytoskeletal analysis. Biochemical approaches such as RhoA activity pull-downs and co-immunoprecipitation of CRMP1/CRMP2 enable mechanistic dissection of DPYSL3 signaling networks. RNA-seq or proteomic profiling can reveal transcriptomic changes upon DPYSL3 disruption. The product is also suited for drug sensitivity screens targeting cytoskeletal pathways and apoptosis assays investigating cell death regulation. For further information or technical support, please contact Ascent Research.
