Optimization of mRNA Synthesis for Cleavage-Free Cas9-Based Epigenetic Editing
발표자
구다영 (서울대학교)
연구책임자
이혁진 (서울대학교)
공동저자
구다영 (서울대학교), 황유나 (서울대학교), 이혁진 (서울대학교)
초록
내용
Conventional genome editing tools, including base editors, enable targeted DNA modification but permanently alter genomic DNA, limiting their use in reversible applications. Epigenetic editing has therefore emerged as a promising alternative, as it allows reversible regulation of gene expression. Since these editors must be transiently expressed and subsequently eliminated, mRNA-based delivery is particularly well suited for this purpose. Here, we optimized the in vitro transcription process to synthesize long epigenetic editing mRNA constructs (>6 kb) encoding dCas9 and epigenetic effectors. Following cellular delivery, temporal protein expression was evaluated by Western blot analysis. The observed time-dependent protein expression profiles suggest a potential relationship between mRNA stability and epigenetic regulatory activity. These results provide a technical foundation for cleavage-free, temporally controlled epigenetic editing using an mRNA-based delivery platform.