The ARID5A Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for the targeted disruption of the human ARID5A gene in the widely used HEK293T host background. This product provides a heterogeneous pool of cells carrying various loss-of-function edits, enabling robust loss-of-function studies without clonal selection. The polyclonal format preserves population-level reproducibility while minimizing the risk of clonal artifacts, making it suitable for high-throughput screening and pooled functional genomics applications.
The host cell line, HEK293T, is a derivative of the human embryonic kidney 293 cell line that stably expresses the SV40 large T antigen, conferring episomal replication of plasmids containing the SV40 origin of replication. This epithelial-like cell line is renowned for its high transfectability and exceptional capacity for viral production, making it a cornerstone model for transient and stable protein expression, lentivirus and retrovirus packaging, and a broad range of biochemical and cell-based assays.
ARID5A (AT-rich interactive domain-containing protein 5A) is a transcription factor that binds AT-rich DNA sequences and plays critical roles in immune cell differentiation, adipogenesis, and osteoblastogenesis. In the IL-6/JAK/STAT3 signaling pathway, STAT3 directly induces ARID5A expression upon cytokine stimulation. ARID5A then promotes Th17 cell differentiation by binding to the il17a promoter and enhancing IL-17A transcription, thereby driving pro-inflammatory gene programs. In mesenchymal lineages, ARID5A interacts with HDAC1, HDAC2, and the SIN3A corepressor complex to modulate the expression of key lineage-determining factors, including PPARG during adipogenesis and RUNX2 during osteoblastogenesis. ARID5A activity is further regulated by TNF-??, STAT5, TGF-??, and BMP2, highlighting its integration into multiple signaling networks.
In the HEK293T background, this ARID5A knockout model provides a versatile platform to dissect the transcription factor??s biochemical and functional properties independent of its immune-specific context. Researchers can reconstitute signaling pathways by ectopically expressing upstream regulators like STAT3 or downstream effectors, and employ luciferase reporter assays to measure ARID5A-dependent promoter activity. Co-immunoprecipitation and ChIP-qPCR can be performed to map protein?Cprotein and protein?CDNA interactions with endogenous or overexpressed binding partners such as HDAC1 and RUNX2. While HEK293T cells are not inherently adipogenic or osteogenic, they can be co-transfected with lineage-specific master regulators (e.g., PPARG or RUNX2) to study ARID5A??s modulatory effects in a controlled manner.
This knockout product supports a wide array of experimental applications, including autoimmune disease modeling (particularly Th17-driven pathologies such as rheumatoid arthritis and multiple sclerosis), adipogenesis and obesity research, osteoblast differentiation studies, and drug target validation. Representative assays include western blotting and RT-qPCR for expression analysis, RNA-seq for transcriptome-wide profiling, flow cytometry for protein-level validation, and differentiation assays when coupled with appropriate induction protocols. For further details or custom requests, please contact Ascent Research.
