The HNRNPD Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population targeting the HNRNPD gene (AUF1) in the human HeLa cervical adenocarcinoma line. This loss-of-function model enables investigation of post-transcriptional gene regulation mediated by HNRNPD/AUF1, which binds AU-rich elements (AREs) in mRNA 3′ untranslated regions and facilitates mRNA decay. The polyclonal format preserves cellular heterogeneity, supporting robust functional analyses.
HeLa cells are an HPV18-positive epithelial cell line derived from cervical adenocarcinoma, widely used in cancer research. They exhibit inactivated p53 and RB, hyper-triploidy, and an epithelial phenotype, recapitulating key features of HPV-driven oncogenesis. This background provides a relevant context for studying cell cycle dysregulation, apoptosis, and drug responses.
HNRNPD/AUF1 is phosphorylated by p38 MAPK, ERK, GSK3??, and PKC in response to stimuli such as oxidative stress and TNF??, regulating its ARE-binding activity. It targets mRNAs encoding Cyclin D1, c-Myc, p21, Bcl-2, COX-2, and GM-CSF for exosome-mediated decay, thus controlling proliferation, survival, and inflammation. HNRNPD interacts with RNA-binding proteins hnRNPA1, KSRP, TTP, and translation factor eIF4G, integrating its function within the p38 MAPK/MK2 and NF-??B/IKK signaling networks.
In HeLa cells, HNRNPD knockout stabilizes ARE-containing transcripts, upregulating proteins like cyclin D1, c-Myc, and Bcl-2. Combined with p53 and RB inactivation, this further disrupts cell cycle checkpoints and apoptotic control, revealing post-transcriptional mechanisms in cervical carcinogenesis. The model is valuable for studying how mRNA turnover cooperates with HPV-mediated oncogenic signaling.
Applications include ARE-luciferase reporter assays, RT-qPCR, RNA-seq, and co-immunoprecipitation with factors such as hnRNPA1 or KSRP. Functional studies employ flow cytometry, viability, and migration assays, while drug sensitivity testing can assess MAPK or NF-??B pathway inhibitors. For further information, contact Ascent Research.
