The ID3 Knockout HGC-27 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HGC-27 human gastric cancer cell line, carrying a targeted disruption of the ID3 gene. This loss-of-function model eliminates expression of the inhibitor of DNA binding 3 (ID3) protein, enabling systematic interrogation of derepressed basic helix-loop-helix (bHLH) transcription factor activity in a gastric carcinoma background. The polyclonal population represents a genetically heterogeneous pool of cells with individual gene-disruption events, offering a robust and versatile resource for functional studies without the confounding influence of single-clone adaptation.
The parental HGC-27 cell line is an adherent epithelial line originally established from a lymph node metastasis of a human gastric carcinoma. It serves as a well-characterized model of invasive gastric cancer, displaying hallmark features of epithelial tumor cells, including robust proliferation, migratory capacity, and tumorigenic potential in xenografts. HGC-27 cells have been extensively employed in gastric cancer research to examine signaling pathways that drive tumor progression, metastasis, and drug resistance, making them an appropriate host for studying ID3-related mechanisms.
ID3 functions as a dominant-negative regulator of class I and II bHLH transcription factors, notably TCF3 (E12/E47), MYOD1, and NEUROD1. By forming inactive heterodimers, ID3 represses transcription of genes involved in cell-cycle arrest and differentiation, such as CDKN1A (p21). ID3 is transcriptionally induced by TGF-?? and BMP ligands via receptor-regulated SMADs: TGF-??1 signals through TGFBR1/2 and SMAD2/3, whereas BMP2 and BMP4 engage BMPR1A/1B and SMAD1/5/8. These pathways converge on ID3 to promote cell proliferation, angiogenesis, and epithelial-mesenchymal transition, and ID3 loss disrupts this hub, derepressing bHLH-mediated transcription of downstream effectors including CCND1, VEGFA, and SNAI1.
In the gastric cancer context, ID3 is frequently overexpressed and correlates with tumor progression and metastasis. Disruption of ID3 in HGC-27 cells eliminates dominant-negative inhibition of bHLH factors, relieving repression of pro-differentiation and anti-proliferative programs. This perturbation is predicted to attenuate cell cycle progression, reduce angiogenic signaling, and impair migratory and invasive capacities. The polyclonal nature of this knockout population provides a genetically heterogeneous loss-of-function model without clonal selection bias.
Typical applications include functional analysis of gastric cancer cell proliferation, apoptosis, and differentiation, as well as dissection of TGF-??/BMP signaling crosstalk. This model supports Western blotting, RT-qPCR, MTS/CCK-8 proliferation assays, colony formation, wound healing, Transwell invasion, Annexin V apoptosis detection, phospho-SMAD profiling, RNA-seq, and immunofluorescence. For further technical inquiries, please contact Ascent Research.
