@article{6183be55d5714490b90fe11a08eba962,
title = "Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection",
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.",
keywords = "flatworm, gene delivery, luminescence, mRNA transfection, planarian, post-transcriptional regulation, Schmidtea mediterranea, somatic transformation",
author = "Hall, {Richard Nelson} and Uri Weill and Leonard Drees and Sergio Leal-Ortiz and Hongquan Li and Margarita Khariton and Chew Chai and Yuan Xue and Benyamin Rosental and Quake, {Stephen R.} and {S{\'a}nchez Alvarado}, Alejandro and Melosh, {Nicholas A.} and Fire, {Andrew Z.} and Rink, {Jochen C.} and Bo Wang",
note = "Funding Information: We thank K. Lei and C. Kuhn for stimulating discussion and E. Liu, D. Burghardt, Y. Lim, and T. Boothe for technical assistance. R.N.H. is supported by an NSF Graduate Research Fellowship. U.W. is supported by an EMBO Long-Term Fellowship. B.W. is a Beckman Young Investigator. U.W. L.D. and J.C.R. are supported by the Max Planck Society. A.Z.F. is supported by an NIH grant (R35GM130366). B.W. and B.R. appreciate the support from an HFSP grant (RGY0085/2019). This work was initiated through a Stanford Bio-X Interdisciplinary Initiative seed grant (IIP9-64) to B.W. A.Z.F. and N.A.M. and later supported by an NSF EDGE grant (IOS-1923534) to B.W. A.S.A. and N.A.M. and a Volkswagen Foundation grant (Az.-99498) to J.C.R. and B.W. Conceptualization, R.N.H. U.W. A.Z.F. J.C.R. and B.W.; methodology, R.N.H. U.W. L.D. S.L.-O. H.L. M.K. and B.R.; investigation, R.N.H. U.W. L.D. C.C. and Y.X.; writing – original draft, R.N.H. J.C.R. and B.W.; writing – review & editing, R.N.H. U.W. S.L.-O. A.S.A. A.Z.F. J.C.R. and B.W. with input from all of the other authors; supervision, S.R.Q. N.A.M. A.Z.F. J.C.R. and B.W.; funding acquisition, A.S.A. N.A.M. A.Z.F. J.C.R. and B.W. S.L.-O. and N.A.M. are co-founders of NAVAN Technologies. Funding Information: We thank K. Lei and C. Kuhn for stimulating discussion and E. Liu, D. Burghardt, Y. Lim, and T. Boothe for technical assistance. R.N.H. is supported by an NSF Graduate Research Fellowship . U.W. is supported by an EMBO Long-Term Fellowship . B.W. is a Beckman Young Investigator . U.W., L.D., and J.C.R. are supported by the Max Planck Society . A.Z.F. is supported by an NIH grant ( R35GM130366 ). B.W. and B.R. appreciate the support from an HFSP grant ( RGY0085/2019 ). This work was initiated through a Stanford Bio-X Interdisciplinary Initiative seed grant (IIP9-64) to B.W., A.Z.F., and N.A.M. and later supported by an NSF EDGE grant ( IOS-1923534 ) to B.W., A.S.A., and N.A.M., and a Volkswagen Foundation grant ( Az.-99498 ) to J.C.R. and B.W. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = oct,
day = "24",
doi = "10.1016/j.crmeth.2022.100298",
language = "English",
volume = "2",
journal = "Cell Reports Methods",
issn = "2667-2375",
publisher = "Cell Press",
number = "10",
}