The DOCK5 Knockout MES-OV Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the MES-OV mouse embryonic stem cell line. This product offers a heterogeneous pool of cells with targeted DOCK5 gene disruptions, enabling loss-of-function studies in a pluripotent context. The polyclonal format provides a convenient model for initial phenotypic screening, reflecting the diversity of CRISPR-mediated edits, and is suitable for high-throughput applications where clonal uniformity is not required.
MES-OV is a mouse embryonic stem cell line from the 129/Sv strain, widely used in pluripotency and developmental research. These cells maintain the ability to differentiate into all three germ layers, providing a versatile platform for studying gene function during early embryogenesis. With robust growth and amenability to genetic manipulation, MES-OV serves as a reliable host for CRISPR/Cas9 editing, and upon directed differentiation, DOCK5 knockout cells can be used to investigate lineage specification, particularly of mesodermal derivatives.
DOCK5 encodes a guanine nucleotide exchange factor that specifically activates Rac and Cdc42 GTPases by facilitating GTP loading. It functions in a complex with ELMO1, which is recruited to the membrane by upstream signals from integrins, PDGFR, and chemokines like SDF-1. Activated Rac1 and Cdc42 engage downstream effectors including PAK1 and LIMK1, leading to cofilin phosphorylation and actin cytoskeleton remodeling. This cascade regulates cell migration, adhesion, and fusion, and is essential for osteoclast-mediated bone resorption.
In MES-OV pluripotent stem cells, DOCK5 knockout allows dissection of Rac/Cdc42-dependent cytoskeletal regulation in self-renewal, early differentiation, and morphogenesis. As DOCK5 is critical for osteoclast function, these cells are valuable for modeling bone remodeling disorders like osteopetrosis when differentiated toward osteoclasts. Additionally, the model reveals impacts on focal adhesion and integrin signaling, processes central to both development and metastasis.
These cells support diverse assays: western blotting for protein depletion, Rac/Cdc42 activation pull-downs to assess signaling, transwell migration assays, and actin immunofluorescence to visualize cytoskeletal defects. Osteoclast differentiation and resorption pit assays allow functional bone biology studies, while RT-qPCR provides transcript-level analysis. For more information, contact Ascent Research.
