The IGF2BP3 Knockout HT29 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal population of HT29 colorectal adenocarcinoma cells with targeted disruption of the IGF2BP3 gene. This polyclonal knockout cell pool is designed for loss-of-function studies of the RNA-binding protein IGF2BP3 in a well-characterized epithelial colorectal cancer background. The polyclonal format provides a heterogeneous mixture of edited alleles, enabling investigation of bulk-knockout phenotypes without cloning artifacts.
The HT29 cell line, derived from a primary human colorectal adenocarcinoma, is a standard model for colorectal cancer research and drug resistance studies. HT29 cells harbor a BRAF V600E mutation and wild-type KRAS, reflecting a clinically significant subtype. These adherent epithelial cells retain key oncogenic signaling dependencies that facilitate investigation of tumorigenesis and therapeutic response.
IGF2BP3 is an m6A reader protein that stabilizes N6-methyladenosine-modified mRNAs and enhances their translation. It operates downstream of transcriptional regulators MYC and ??-catenin/TCF, with inputs from Wnt and TGF-?? pathways. IGF2BP3 interacts with METTL3, YTHDF2, HNRNPA2B1, and the RNA methyltransferase complex, and it targets oncogenic transcripts including CD44, MYC, IGF2, HMGA2, SNAI2, and CCND1. This post-transcriptional regulon amplifies protein expression that drives proliferation, migration, and stemness.
In the HT29 colorectal cancer model, IGF2BP3 likely supports malignant phenotypes driven by BRAF V600E and aberrant Wnt/??-catenin signaling. Disruption of IGF2BP3 allows dissection of how m6A-dependent mRNA stabilization contributes to tumor growth, epithelial-mesenchymal transition, and drug resistance. This knockout population provides a genetic tool to study the gene’s role in maintaining oncogenic mRNA translation in a relevant colorectal cancer background.
Applications include research on post-transcriptional gene regulation, cancer epitranscriptomics, and oncogenic signaling. Typical assays are RNA immunoprecipitation, RT-qPCR, Western blotting, m6A-RIP sequencing, luciferase reporter assays, colony formation, migration/invasion assays, immunofluorescence, and polysome profiling. These methods enable analysis of mRNA half-life, translational control, and pathway activity following IGF2BP3 loss. The model supports drug-response studies and functional genomics screens. For additional technical details, contact Ascent Research.
