A simplified method to produce mRNAs and functional proteins from synthetic double-stranded DNA templates

Biotechniques. 2020 Oct;69(4):281-288. doi: 10.2144/btn-2020-0037. Epub 2020 Aug 20.

Abstract

We present a method to synthesize mRNAs from synthetic DNA templates that produce biologically active proteins. To illustrate utility, we constructed five unique synthetic DNA templates, produced mRNAs and demonstrated biologic activity of their translated proteins. Examples include secreted luciferase, enhanced green fluorescence protein, IL-4, and IL-12A and IL-12B to form active IL-12. We propose that this method offers a cost- and time-saving alternative to plasmid-based cloning.

Keywords: alternative to molecular cloning; cell transfection; chemical DNA synthesis; protein translation; synthetic mRNA from DNA template.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cloning, Molecular
  • DNA / biosynthesis
  • DNA / genetics*
  • Green Fluorescent Proteins / chemistry
  • Interleukin-12 / biosynthesis
  • Interleukin-12 / genetics
  • Interleukin-4 / biosynthesis
  • Interleukin-4 / genetics
  • Luciferases / genetics
  • Plasmids / genetics
  • Protein Biosynthesis / genetics*
  • Proteins / genetics*
  • RNA, Messenger / biosynthesis*
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • Templates, Genetic

Substances

  • Proteins
  • RNA, Messenger
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Interleukin-12
  • Interleukin-4
  • DNA
  • Luciferases