The ID3 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the ID3 gene. This product consists of a heterogeneous pool of cells carrying diverse CRISPR/Cas9-mediated disruptions at the ID3 locus, providing a robust model to interrogate ID3-dependent biological processes without the selection of a single clonal isolate. The polyclonal format mitigates clonal artifacts and offers a representative knockout background for functional genomics, signaling pathway dissection, and phenotypic screening applications.
The parental HEK293T cell line is a widely utilized human embryonic kidney epithelial model that stably expresses the SV40 large T antigen, enabling high-level plasmid amplification and efficient protein production. These cells are a mainstay in mammalian cell biology for recombinant protein expression, lentiviral and retroviral packaging, and transient or stable transgene studies. Their robust growth characteristics, ease of transfection, and well-characterized signaling landscape render them an optimal chassis for generating gene-edited derivatives to explore diverse molecular mechanisms.
ID3 functions as a dominant-negative inhibitor of basic helix-loop-helix (bHLH) transcription factors by forming transcriptionally inactive heterodimers with E-proteins, including TCF3, TCF4, and TCF12, thereby preventing their binding to E-box regulatory elements and repressing target genes involved in differentiation and cell cycle arrest. Its expression is transcriptionally induced by upstream regulators such as TGF-??, BMPs, Notch ligands, HIF1??, E2F, Myc, and Smad proteins, linking ID3 to multiple signaling cascades. Downstream, ID3 suppresses the activity of myogenic and neurogenic factors like MyoD and neurogenin, and modulates cell cycle regulators p21 (CDKN1A) and p16 (CDKN2A). ID3 also engages in cross-talk with other ID family members (ID1, ID2, ID4) and integrates signals from receptors including TGFBR1/2, BMPR1/2, and NOTCH1-4, with pathway mediators such as SMAD2/3/4, SMAD1/5/8, HEY1, and HES1.
In the HEK293T background, ID3 knockout provides a pertinent system to study how disruption of bHLH inhibition impacts cellular programs in an epithelial context with intact TGF-??/BMP/Notch signaling machinery. Because HEK293T cells exhibit a partially transformed phenotype and express SV40 large T antigen, which affects cell cycle regulation, the loss of ID3 can reveal synergistic or compensatory interactions with these pathways. This model allows researchers to dissect ID3’s role in governing proliferation, apoptosis, and epithelial-to-mesenchymal transition-related processes, and to evaluate signaling crosstalk in a genetically accessible and highly transfectable host.
These polyclonal knockout cells are suited for a broad array of experimental applications, including western blotting and RT-qPCR to confirm ID3 ablation and monitor downstream targets, RNA-seq for transcriptome-wide profiling of ID3-dependent gene networks, and functional assays such as MTT/BrdU proliferation, Annexin V apoptosis, and migration/invasion studies. They facilitate drug target validation and screening for small molecules that modulate ID3-regulated pathways. Additionally, these cells support reporter gene assays for E-box activity and immunofluorescence-based localization studies. For further information, please contact Ascent Research.
