The ID3 Knockout LoVo Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the LoVo colorectal adenocarcinoma cell line, providing a loss-of-function model for inhibitor of DNA binding 3 (ID3). This product consists of a heterogeneous pool of cells carrying targeted disruptions of the ID3 gene, enabling functional genomics studies without clonal selection. The polyclonal format avoids single-clone artifacts and is well-suited for analyzing gene-dependent phenotypes in cancer progression.
The host LoVo line originates from a supraclavicular lymph node metastasis of a 56-year-old male colon adenocarcinoma patient. These adherent epithelial cells are a classic model of human colon cancer, characterized by aberrant Wnt/??-catenin and TGF-?? signaling, and are extensively used to investigate tumorigenesis, epithelial-mesenchymal transition (EMT), and metastatic dissemination.
ID3 encodes a dominant-negative helix-loop-helix protein that inhibits bHLH transcription factors by forming inactive heterodimers, thereby blocking target gene activation. In LoVo cells, ID3 expression is driven by the ??-catenin/TCF complex and TGF-??/SMAD pathways, integrating signals from Notch (NICD), c-MYC, and EGF. ID3 represses p21/WAF1 and E-cadherin transcription, while upregulating MMP2, MMP9, Cyclin D1, N-cadherin, and BCL2, promoting proliferation, EMT, and invasion. It interacts with E-proteins, SMAD3, and ID family members, forming a complex regulatory network.
Disruption of ID3 in this colorectal cancer model attenuates Wnt/??-catenin and TGF-?? signaling, leading to reduced invasive capacity and restored expression of cell-cycle inhibitors. This polyclonal knockout system thus enables detailed dissection of ID3??s role in sustaining malignant phenotypes, including EMT and matrix remodeling. It serves as a powerful tool for target validation and drug screening efforts in colon cancer.
Applications include Western blot and RT-qPCR to verify ID3 loss and downstream target modulation, Transwell assays for migration and invasion, and luciferase reporters for Wnt pathway activity. Proliferation and apoptosis can be assessed by MTT assay and flow cytometry, respectively. RNA-seq provides transcriptome-wide insights, while immunofluorescence visualizes EMT marker redistribution. For further information, please contact Ascent Research.
