Abstract

mRNA medicines can be used to express therapeutic proteins, but the production of such proteins in non-target cells has a risk of adverse effects. To accurately distinguish between therapeutic target and nontarget cells, it is desirable to utilize multiple proteins expressed in each cell as indicators. To achieve such multi-input translational regulation of mRNA medicines, in this study, we engineered Rhodothermus marinus (Rma) DnaB intein to develop “caged Rma DnaB intein” that enables conditional reconstitution of full-length translational regulator protein from split fragments. By combining the caged Rma DnaB intein, the split translational regulator protein, and target protein-binding domains, we succeeded in target protein-dependent translational repression of mRNA in human cells. In addition, the caged Rma intein showed orthogonality to the previously reported Nostoc punctiforme (Npu) DnaE-based caged intein. Finally, by combining these two orthogonal caged inteins, we developed an mRNA-based logic gate that regulates translation based on the expression of multiple intracellular proteins. This study provides important information to develop safer mRNA medicines.

Details

Title
Development of a new caged intein for multi-input conditional translation of synthetic mRNA
Author
Yang, Tingting 1 ; Nakanishi, Hideyuki 2 ; Itaka, Keiji 2 

 Tokyo Medical and Dental University (TMDU), Department of Biofunction Research, Institute of Biomaterials and Bioengineering, Tokyo, Japan (GRID:grid.265073.5) (ISNI:0000 0001 1014 9130) 
 Tokyo Medical and Dental University (TMDU), Department of Biofunction Research, Institute of Biomaterials and Bioengineering, Tokyo, Japan (GRID:grid.265073.5) (ISNI:0000 0001 1014 9130); Osaka University, Center for Infectious Disease Education and Research (CiDER), Osaka, Japan (GRID:grid.136593.b) (ISNI:0000 0004 0373 3971) 
Pages
9988
Publication year
2024
Publication date
2024
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
3049547944
Copyright
© The Author(s) 2024. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.