Description

The SERPINE1 Knockout DF1 Cell Line is a CRISPR/Cas9-edited knockout cell line engineered from the DF1 chicken embryo fibroblast line. This model disrupts the SERPINE1 gene, which encodes plasminogen activator inhibitor-1 (PAI-1), an essential serpin that regulates fibrinolysis and extracellular matrix (ECM) homeostasis. The cell line provides a validated platform for loss-of-function studies in avian cell biology.

DF1 is a spontaneously immortalized fibroblast line derived from the East Lansing Line (ELL-0) chicken embryo. These cells are fully permissive for avian virus replication and highly transfectable, making them a standard host for virological research, recombinant protein production, and investigation of fibroblast-specific functions. Their embryonic origin and stable phenotype ensure reproducible experiments across passages.

PAI-1 functions by irreversibly inhibiting urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA), thereby controlling plasmin-mediated ECM degradation and cell motility. PAI-1 expression is transcriptionally regulated by TGF-??1, TNF, IL-1??, HIF1A, glucocorticoids, insulin, and p53. At the protein level, PAI-1 binds vitronectin, uPA/tPA complexes, LRP1, and integrins, facilitating internalization and signal transduction. Disruption of SERPINE1 abolishes this inhibitory function, resulting in unchecked uPA and tPA activity, elevated plasmin levels, and activation of matrix metalloproteinases such as MMP2 and MMP9, which collectively promote ECM remodeling and alter cell adhesion and migration.

In DF1 fibroblasts, SERPINE1 knockout disturbs the proteolytic balance critical for fibrinolysis and tissue remodeling. This cell line is particularly suited for dissecting TGF-??/SMAD2/3 signaling and its crosstalk with plasmin-dependent pathways. The spontaneously immortalized nature of DF1 permits indefinite culture while retaining key fibroblast traits, enabling consistent studies of wound healing, senescence-associated secretory phenotype, and viral replication where host protease inhibitors like PAI-1 may influence infection dynamics.

Researchers can employ this knockout cell line in a variety of assays: Western blot and RT-qPCR for PAI-1 validation, plasminogen activator zymography to assess uPA/tPA activity, scratch wound and transwell assays for migration/invasion, and fluorescence-based ECM degradation assays. It is useful for modeling thrombosis, fibrosis, cancer metastasis, cardiovascular disease, metabolic syndrome, and preeclampsia in an avian context, as well as for optimizing avian virus production. For product orders or technical inquiries, please contact Ascent Research.