Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection

通过体细胞 mRNA 转染在涡虫 Schmidtea mediterranea 中进行异源报告基因表达

阅读:9
作者:Richard Nelson Hall, Uri Weill, Leonard Drees, Sergio Leal-Ortiz, Hongquan Li, Margarita Khariton, Chew Chai, Yuan Xue, Benyamin Rosental, Stephen R Quake, Alejandro Sánchez Alvarado, Nicholas A Melosh, Andrew Z Fire, Jochen C Rink, Bo Wang

Abstract

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.

特别声明

1、本页面内容包含部分的内容是基于公开信息的合理引用;引用内容仅为补充信息,不代表本站立场。

2、若认为本页面引用内容涉及侵权,请及时与本站联系,我们将第一时间处理。

3、其他媒体/个人如需使用本页面原创内容,需注明“来源:[生知库]”并获得授权;使用引用内容的,需自行联系原作者获得许可。

4、投稿及合作请联系:info@biocloudy.com。