A Recombinant Collagen-mRNA Platform for Controllable Protein Synthesis

Chembiochem. 2015 Jul 6;16(10):1415-9. doi: 10.1002/cbic.201500205. Epub 2015 May 26.

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.

Keywords: collagen; mRNA; protein synthesis; ribosomes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell-Free System* / metabolism
  • Collagen / chemistry
  • Collagen / genetics*
  • Escherichia coli / genetics
  • Green Fluorescent Proteins / analysis
  • Green Fluorescent Proteins / genetics
  • Luciferases / analysis
  • Luciferases / genetics
  • Luminescent Agents / analysis
  • Luminescent Agents / metabolism
  • Maltose-Binding Proteins / chemistry
  • Maltose-Binding Proteins / genetics
  • Protein Biosynthesis*
  • Protein Multimerization
  • Protein Stability
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics

Substances

  • Luminescent Agents
  • Maltose-Binding Proteins
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Collagen
  • Luciferases