The ACTA2 Knockout 786-O Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ACTA2 gene, which encodes alpha-smooth muscle actin (??-SMA). This product provides a heterogeneous pool of 786-O renal carcinoma cells with disrupted ACTA2, enabling robust loss-of-function analyses without clonal selection bias. The polyclonal format preserves biological variability and is ideal for population-level phenotypic screening. Abrogation of ??-SMA expression in these cells offers a direct means to study cytoskeletal-dependent processes.
The parental 786-O line is a human clear cell renal cell adenocarcinoma model with biallelic VHL inactivation, leading to constitutive HIF stabilization. It is widely used to investigate renal cancer biology, hypoxia response, and tumor progression. The cells display typical epithelial morphology, providing a relevant context for examining epithelial-mesenchymal transition (EMT) and metastatic mechanisms.
??-SMA is a key actin isoform forming contractile stress fibers, essential for cell contractility, migration, and ECM remodeling. Transcription of ACTA2 is primarily activated by TGF??1 through SMAD2/3 and the co-factors MRTF-A and SRF, with modulation by PDGF, YAP1, and Rho/ROCK signaling. ??-SMA interacts with myosin, tropomyosin, and caldesmon to generate force, and its disruption impairs TGF??-induced cytoskeletal reorganization and downstream contractile functions.
In 786-O cells, ACTA2 knockout abolishes ??-SMA protein and prevents stress fiber assembly, blocking the myofibroblastic phenotype typically triggered by TGF??1. This attenuation of mesenchymal traits, such as elongated morphology and increased motility, makes the model valuable for dissecting the role of ??-SMA in renal cancer cell plasticity and invasion. It allows precise assessment of how ??-SMA contributes to metastatic behavior.
Typical applications include investigating TGF??-driven EMT, screening anti-fibrotic or anti-metastatic compounds, and analyzing actin dynamics. Common assays involve western blotting for ACTA2 and EMT markers, immunofluorescence for F-actin, scratch wound and Transwell migration/invasion assays, and RT-qPCR for EMT transcription factors following TGF??1 treatment. These applications leverage the knockout to link molecular signaling to functional outcomes. For further information, contact Ascent Research.
