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

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.

Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR

“…Next, we tested whether hid-RT-PCR could be improved by addition of the chemical, thermostable, RNase inhibitor polyvinylsulfonic acid (PVSA), and/or the chelating agent ethylenediaminetetraacetic acid (EDTA) during heat inactivation. PVSA halves in vitro RNase A activity (50% inhibition, IC 50 ) at the concentration 150 μg/ml and halves the RNase activity of E. coli lysate (IC 50 ) at 430 μg/ml 12 . At the same time, PVSA might inhibit RT-PCR.…”

Identification of an optimal method for extracting RNA from human skin biopsy, using domestic pig as a model system

“…Thus, obtaining highly degraded RNA from enzymatically treated samples was an expected result. However, if, for some reason, enzymatic digestion at 37 °C is needed, RNase inhibitors are strongly recommended 30,31 .…”

COVID-19: A challenge for electrochemical biosensors

“…In the case of possible nucleases within the sample, the addition of ethylenediamine-tetraacetic acid (EDTA) or glycol-bis(2-aminoethylether)-N,N,N,N-tetraacetic acid (EGTA) is recommended to chelate divalent and trivalent cations required as cofactors for several nucleases. However, neither EDTA nor EGTA inactivates RNases hence alternative approaches such as the use of diethyl pyrocarbonate (DEPC)-treated or polyvinyl sulfonic acid-treated solutions, have been developed [ 68 , 69 ]. Although such chemical treatment of samples can interfere with subsequent PCR amplification, hybridization of nucleic acids and application of electrochemical biosensors should not be influenced.…”