Therapeutic Silencing and Knockout of the RF Gene via Combined Genetic, Epigenetic, and siRNA Mechanisms
Year: 2025
Type: preprint
Abstract: The RF gene has been implicated in critical pathogenic and cellular processes, making it a promising target for precision therapeutics. In this study, we present a comprehensive approach integrating genetic knockout, epigenetic modulation, and small interfering RNA (siRNA) silencing to achieve robust and sustained inhibition of RF gene expression. CRISPR/Cas9-mediated gene knockout was employed to induce permanent loss of function at the genomic level, while targeted epigenetic editing using CRISPR/dCas9 fused to histone-modifying enzymes facilitated chromatin remodeling and transcriptional repression. In parallel, chemically modified siRNAs were designed to promote rapid, transient degradation of RF mRNA, enabling immediate phenotypic modulation prior to complete genetic inactivation. Functional assays demonstrated significant reductions in RF gene transcription and translation, accompanied by downstream suppression of pathogenic signaling pathways. This multimodal strategy offers a synergistic platform capable of both rapid and enduring gene silencing, highlighting its potential for therapeutic applications in diseases driven by aberrant RF gene activity.
Author: Saif Yaseen Hasan
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
OpenAlex ID: W4413549443