The EAF2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the EAF2 gene in human HEK293T cells. This polyclonal population provides a mixed genetic background, enabling robust loss-of-function studies while representing the heterogeneous editing outcomes typical of CRISPR/Cas9-mediated gene disruption. The product is suitable for analyzing EAF2-dependent cellular processes without the need for single-cell cloning, maintaining biological variability relevant to population-level experiments.
HEK293T cells are a widely used human embryonic kidney epithelial cell line that stably expresses the SV40 large T antigen. This antigen enhances episomal replication of plasmids containing the SV40 origin, making HEK293T cells highly efficient for transient protein expression and viral vector production. The cells exhibit adherent epithelial morphology and are commonly employed in molecular and cellular biology laboratories for recombinant protein production, lentivirus packaging, and transcriptional studies.
EAF2 encodes a transcriptional elongation factor that functions as a core component of the super elongation complex (SEC), interacting directly with ELL and RNA polymerase II to promote efficient transcriptional elongation. EAF2 is also a modulator of androgen receptor (AR) signaling; it interacts with the AR and influences the expression of downstream target genes such as KLK3 and TMPRSS2. Mechanistically, EAF2 acts downstream of androgen-bound AR and is part of a network that includes AFF4, CDK9, and Cyclin T1 within the SEC. In prostate cancer, EAF2 expression is frequently downregulated, and its loss is associated with enhanced AR-driven proliferation, supporting its role as a potential tumor suppressor.
Knockout of EAF2 in the HEK293T background provides a reductionist epithelial model to dissect the molecular functions of EAF2 independent of tissue-specific factors. The SV40 large T antigen present in these cells may interact with transcriptional regulators, creating a context in which EAF2’s role in elongation and AR signaling can be assessed with high transfection efficiency and ease of manipulation. This model enables the study of how EAF2 loss affects RNA polymerase II processivity and AR-mediated gene expression programs in a well-characterized cellular environment, facilitating insights into its tumor-suppressive mechanisms.
These polyclonal knockout cells are ideal for a range of functional genomics applications, including genome-wide transcriptomic profiling via RNA-seq to identify EAF2-dependent gene networks, quantitative RT-qPCR and Western blotting to validate target gene expression changes, and co-immunoprecipitation assays to examine altered protein-protein interactions within the super elongation complex. They also support androgen receptor reporter assays to measure AR transcriptional activity and proliferation assays to evaluate the growth-suppressive functions of EAF2. For additional technical specifications or custom inquiries, please contact Ascent Research.
