Polyvinylsulfonic acid: A Low-cost RNase inhibitor for enhanced RNA preservation and cell-free protein translation

Bioengineered. 2018 Jan 1;9(1):90-97. doi: 10.1080/21655979.2017.1313648. Epub 2017 Jun 29.

Abstract

The effectiveness and economics of polyvinyl sulfonic acid (PVSA) as a ribonuclease inhibitor for in vitro systems is reported. PVSA was shown to inhibit RNA cleavage in the presence of RNase A as well as in the presence of Escherichia coli lysate, suggesting that PVSA can act as a broader ribonuclease inhibitor. In addition, PVSA was shown to improve the integrity of mRNA transcripts by up to 5-fold in vitro as measured by their translational viability. Improved preservation of mRNA transcripts in the presence of PVSA under common RNA storage conditions is also reported. A cost comparison with commercially available RNAse inhibitors indicates the economic practicality of PVSA which is approximately 1,700 times less expensive than commonly used ribonuclease inhibitors. PVSA can also be separated from RNA by alcohol precipitation for applications that may be sensitive to the presence of PVSA.

Keywords: RNA storage; RNase inhibitor; in vitro transcription and translation; nuclease inhibition; polyvinyl sulfonic acid.

MeSH terms

  • Bacterial Proteins / antagonists & inhibitors*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Enzyme Assays
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / economics
  • Enzyme Inhibitors / pharmacology*
  • Escherichia coli / enzymology
  • Escherichia coli / genetics
  • Kinetics
  • Polyvinyls / chemistry
  • Polyvinyls / economics
  • Polyvinyls / pharmacology*
  • Protein Biosynthesis*
  • RNA Stability
  • RNA, Messenger / chemistry
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Ribonucleases / antagonists & inhibitors*
  • Ribonucleases / genetics
  • Ribonucleases / metabolism
  • Subcellular Fractions / metabolism
  • Sulfonic Acids / chemistry
  • Sulfonic Acids / economics
  • Sulfonic Acids / pharmacology*
  • Transcription, Genetic

Substances

  • Bacterial Proteins
  • Enzyme Inhibitors
  • Polyvinyls
  • RNA, Messenger
  • Sulfonic Acids
  • lyapolate
  • Ribonucleases