The EID3 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney cell line. This product provides a loss-of-function model for targeted disruption of the EID3 gene, eliminating endogenous EID3 protein expression across a heterogeneous cell pool. By employing a population-level knockout strategy, researchers can investigate gene function without the biases associated with monoclonal selection, making it ideal for robust, reproducible functional genomics studies. The polyclonal format ensures that observed phenotypes reflect the average behavior of multiple independent editing events, offering a realistic assessment of pathway modulation in a mammalian cell context.
The HEK293T host cell line is a human embryonic kidney epithelial line stably expressing the SV40 large T antigen and adenovirus E1A proteins, which confer high transfection efficiency and episomal plasmid amplification. This background is particularly advantageous for transient and stable expression studies, protein production, and lentiviral packaging. The cells grow rapidly in standard culture conditions and are well-characterized, facilitating seamless integration into existing experimental workflows. Their robust nature supports a broad range of downstream applications, including transcriptomic, proteomic, and cell-based functional assays, making them a versatile chassis for gene-edited model generation.
EID3 encodes a transcriptional corepressor that directly interacts with the histone acetyltransferases EP300 and CREBBP, inhibiting their catalytic activity and thereby reducing histone H3 acetylation at target gene promoters. This repression is critical for negative regulation of MYOD-dependent transcription, linking EID3 to control of muscle differentiation and broader cell fate decisions. Upstream, EID3 expression is responsive to the NOTCH signaling pathway and the MYOD transcription factor itself, creating a regulatory feedback loop that fine-tunes gene expression programs governing proliferation and differentiation. Thus, EID3 sits at the nexus of epigenetic machinery and developmental signaling, orchestrating chromatin states essential for cellular identity.
In the HEK293T context, knockout of EID3 is predicted to relieve transcriptional repression at MYOD target loci, leading to increased histone acetylation and altered expression of cell cycle regulators. This model enables precise dissection of how EID3 modulates EP300/CREBBP activity without confounding endogenous background signals. The polyclonal population format ensures that the functional consequences of EID3 loss are assessed across a spectrum of genetic perturbations, capturing a more complete picture of its role in chromatin remodeling and gene regulation. This is especially valuable for studying pathways that intersect with embryogenesis and oncogenic transformation, where subtle changes in transcriptional control can have profound effects.
Detailed applications of these knockout cells include transcriptome-wide RNA sequencing to map EID3-dependent gene networks, chromatin immunoprecipitation coupled with qPCR (ChIP-qPCR) to evaluate histone H3 acetylation changes at EP300/CREBBP target sites, and co-immunoprecipitation assays to probe EP300-CREBBP complex integrity. Additionally, cell cycle analysis by flow cytometry, proliferation assays, and MYOD-responsive luciferase reporter assays can delineate functional outcomes of EID3 disruption. The cells are also suitable for high-content imaging to monitor differentiation markers and for small molecule screens aimed at modulating EP300/CREBBP activity. For further technical details, please contact Ascent Research.
