The PWARSN Knockout HK-2 Cell Line is a CRISPR/Cas9-edited human kidney proximal tubule epithelial cell line featuring targeted disruption of the PWARSN gene. This loss-of-function model eliminates expression of the PWARSN long non-coding RNA, which serves as a host gene for multiple copies of the SNORD116 small nucleolar RNAs. By employing CRISPR/Cas9-mediated gene disruption, the cell line provides a stable and renewable resource for investigating the molecular functions of PWARSN and its downstream snoRNA effectors in a renal epithelial context, without specification of the precise editing pattern or clonal origin.
The parental HK-2 cell line is an immortalized proximal tubule epithelial cell line derived from normal adult human kidney. HK-2 cells are widely used to study renal reabsorption, secretion, and electrolyte and fluid balance, retaining many characteristics of primary proximal tubular epithelium, including transporter expression and metabolic activities. This normal-derived background avoids the oncogenic mutations present in cancer kidney lines, making it particularly suitable for modeling physiological renal epithelial cell biology and stress responses.
PWARSN functions molecularly as a long non-coding RNA subject to imprinting control, regulated by DNA methylation and the imprinting center, with interaction with the insulator protein CTCF. It acts upstream of SNORD116 snoRNAs, which associate with Fibrillarin and NOP56 within the snoRNP complex to guide 2′-O-methylation of ribosomal RNAs. Disruption of PWARSN abolishes SNORD116 expression, leading to loss of rRNA methylation and potential dysregulation of ribosome biogenesis and translation. This pathway may intersect with mTOR signaling and influence cellular stress responses in kidney epithelial cells.
In the context of immortalized proximal tubule epithelial cells, the PWARSN knockout model holds particular significance for dissecting the role of imprinted lncRNAs in renal homeostasis. Proximal tubule cells exhibit high metabolic demand and rely on robust protein synthesis; loss of SNORD116-mediated rRNA modifications could perturb ribosome function and translational control, offering a unique tool to explore whether imprinting dysregulation contributes to renal dysfunction or stress signaling, areas less studied compared to the neuronal pathologies of Prader-Willi and Angelman syndromes.
This knockout cell line is well-suited for a range of research applications, including RT-qPCR validation of PWARSN and SNORD116 expression, RNA-seq to profile transcriptome-wide changes, and ribosomal RNA modification analysis to confirm loss of 2′-O-methylation. Western blotting for ribosomal proteins and proliferation assays enable functional assessments of translational impacts. It supports investigation of lncRNA-mediated imprinting in renal biology, functional characterization of SNORD116 in epithelial cells, and drug screening for Prader-Willi syndrome phenotypes. For further information, please contact Ascent Research.
