Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
Department of Tissue Dynamics and Regeneration, Max Planck Institute for Multidisciplinary Sciences, Göttingen 37077, Germany.
Cell Rep Methods. 2022 Sep 20;2(10):100298. doi: 10.1016/j.crmeth.2022.100298. eCollection 2022 Oct 24.
Planarians have long been studied for their regenerative abilities. Moving forward, tools for ectopic expression of non-native proteins will be of substantial value. Using a luminescent reporter to overcome the strong autofluorescence of planarian tissues, we demonstrate heterologous protein expression in planarian cells and live animals. Our approach is based on the introduction of mRNA through several nanotechnological and chemical transfection methods. We improve reporter expression by altering untranslated region (UTR) sequences and codon bias, facilitating the measurement of expression kinetics in both isolated cells and whole planarians using luminescence imaging We also examine protein expression as a function of variations in the UTRs of delivered mRNA, demonstrating a framework to investigate gene regulation at the post-transcriptional level. Together, these advances expand the toolbox for the mechanistic analysis of planarian biology and establish a foundation for the development and expansion of transgenic techniques in this unique model system.
涡虫一直因其再生能力而受到研究。展望未来，异位表达非天然蛋白的工具将具有重要价值。我们使用发光报告基因来克服涡虫组织的强烈自发荧光，从而在涡虫细胞和活体动物中证明了异源蛋白的表达。我们的方法基于通过几种纳米技术和化学转染方法引入 mRNA。我们通过改变非翻译区 (UTR) 序列和密码子偏爱性来提高报告基因的表达，从而使用发光成像法在分离细胞和整个涡虫中方便地测量表达动力学。我们还研究了作为递达 mRNA 的 UTR 变化的函数的蛋白质表达，展示了一种用于在转录后水平研究基因调控的框架。总之，这些进展扩展了涡虫生物学的机制分析工具包，并为在这个独特的模型系统中开发和扩展转基因技术奠定了基础。
