A Recombinant Collagen-mRNA Platform for Controllable Protein Synthesis.

Liping Sun,Miri Krupkin,Tamar Unger,Donna Matzov,Hagith Yonath,Ella Zimmerman,Shirley Shulman Daube,Ada Yonath,Anat Bashan,Shira Albeck,Yunjing Xiong,Yoav Peleg

doi:10.1002/cbic.201500205

Abstract

We have developed a collagen–mRNA platform for controllable protein production that is intended to be less prone to the problems associated with commonly used mRNA therapy as well as with collagen skin-healing procedures. A collagen mimic was constructed according to a recombinant method and was used as scaffold for translating mRNA chains into proteins. Cysteines were genetically inserted into the collagen chain at positions allowing efficient ribosome translation activity while minimizing mRNA misfolding and degradation. Enhanced green fluorescence protein (eGFP) mRNA bound to collagen was successfully translated by cell-free Escherichia coli ribosomes. This system enabled an accurate control of specific protein synthesis by monitoring expression time and level. Luciferase–mRNA was also translated on collagen scaffold by eukaryotic cell extracts. Thus we have demonstrated the feasibility of controllable protein synthesis on collagen scaffolds by ribosomal machinery.

Highlights

We have developed a collagen–messenger RNA (mRNA) platform for controllable protein production that is intended to be less prone to the problems associated with commonly used mRNA therapy as well as with collagen skin-healing procedures
Enhanced green fluorescence protein mRNA bound to collagen was successfully translated by cell-free Escherichia coli ribosomes
Nanoparticles encapsulating luciferase-encoding plasmid and Escherichia coli extract are capable of autonomous synthesis of protein in vitro and in vivo.[4] mRNA vectors have been used as templates for protein synthesis rather than DNA plasmid due to RNA’s low toxicity, low immunogenicity, and high expression efficacy in vivo

Summary

Introduction

We have developed a collagen–mRNA platform for controllable protein production that is intended to be less prone to the problems associated with commonly used mRNA therapy as well as with collagen skin-healing procedures. A Recombinant Collagen–mRNA Platform for Controllable Protein Synthesis Liping Sun,[a] Yunjing Xiong,[a] Anat Bashan,[b] Ella Zimmerman,[b] Shirley Shulman Daube,[b] Yoav Peleg,[b] Shira Albeck,[b] Tamar Unger,[b] Hagith Yonath,[c, d] Miri Krupkin,[b] Donna Matzov,[b] and Ada Yonath*[b]

Results

Conclusion