Lrpprc Knockout B16-F10 Cell Line

The Lrpprc Knockout B16-F10 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the B16-F10 mouse melanoma line, with targeted disruption of the Lrpprc gene. It provides a stable loss-of-function model for investigating Lrpprc, a scaffold protein that recruits phosphatases to focal adhesions. This cell line eliminates Lrpprc protein function, allowing direct assessment of its role in adhesion signaling without the limitations of pharmacological or transient methods. The cell line is maintained under standard culture conditions and can be used directly in migration and adhesion assays.

B16-F10 is a highly metastatic subline of the C57BL/6 B16 melanoma, widely used for studying tumor metastasis and cell motility. Its aggressive migration and invasion, coupled with a syngeneic background, facilitate in vivo transplantation and in vitro assays of adhesion dynamics and Rho GTPase-driven cytoskeletal reorganization. B16-F10 cells are characterized by elevated expression of integrins and active Rho GTPases, which drive their motility. The Lrpprc knockout in this context allows dissection of the specific contribution of phosphatase scaffolding to these dynamic processes. The knockout line in this background ensures sensitive detection of Lrpprc-dependent metastatic phenotypes.

Lrpprc acts as a molecular scaffold, recruiting PTPN12 and other phosphatases to focal adhesions, where it dephosphorylates SRC family kinases, FAK, and paxillin downstream of integrins and growth factor receptors (PDGFR, EGFR). It interacts with p130Cas and actin regulators to modulate Rho GTPases (RAC1, ROCK) and MAPK/ERK signaling, controlling adhesion turnover and migration. This dephosphorylation activity terminates signaling events initiated by integrin clustering and growth factor binding, making Lrpprc a key negative regulator of adhesion complex turnover. Consequently, its loss disrupts the balance between adhesion assembly and disassembly, leading to hyperphosphorylation of adhesion complex components.

In the B16-F10 melanoma model, Lrpprc knockout is expected to impair focal adhesion disassembly, reducing migration and invasion through extracellular matrix. Because B16-F10 cells rely on rapid adhesion turnover for efficient invasion, the knockout cell line may exhibit reduced metastatic ability, providing a tool to study the adhesion-related checkpoints in metastasis. This cell line provides a genetic system to dissect how phosphatase scaffolding at adhesions influences melanoma progression, specifically evaluating nodes within the integrin?CRho GTPase?CMAPK axis critical for metastasis.

Applications include wound healing and transwell migration/invasion assays to measure 2D motility and matrix penetration, immunofluorescence for focal adhesion proteins (paxillin, vinculin), and phospho-western blotting for FAK (pY397) and SRC (pY416). GTPase activity assays for RAC1 and RhoA connect phenotypic changes to signaling outputs. These approaches enable comprehensive analysis of the interplay between Lrpprc, adhesion dynamics, and metastatic signaling. The model suits mechanistic metastasis studies, functional genomics screens, and anti-metastatic compound testing. For further information, contact Ascent Research.