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

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

Planarians have long been studied for their regenerative abilities, but they possess limited genetic tools due to challenges in gene delivery, expression, and detection, despite decades of work. We developed a toolbox for heterologous protein expression in planarian cells and in live animals. Specifically, we identified and optimized nanotechnological and chemical transfection methods to efficiently deliver mRNA encoding nanoluciferase into somatic cells, including planarian adult stem cells (neoblasts). The use of a luminescent reporter allowed us to quantitatively measure protein expression through spectroscopy and microscopy, thus overcoming the strong autofluorescent background of planarian tissues. Using this platform, we investigated the use of endogenous untranslated region (UTR) sequences and codon usage bias to post-transcriptionally alter gene expression. Our work provides a strong foundation for advancing exogenous gene expression and for the rapid prototyping of genetic constructs to accelerate the development of transgenic techniques in planarians.