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Suppression of mRNA Nanoparticle Transfection in Human Fibroblasts by Selected Interferon Inhibiting Small Molecule

Yang Liu1, Manoj N Krishnan2, Kyle K L Phua3

  • 1Department of Chemical and Biomolecular Engineering, Faculty of Engineering, National University of Singapore, 1 Engineering Drive 3, Singapore 117580, Singapore. johnliuyang@nus.edu.sg.

Biomolecules
|August 1, 2017
PubMed

Insights

Small molecules, including interferon inhibitors, were tested to improve in vitro transcribed messenger RNA (IVT mRNA) transfection in human fibroblasts. The study found no enhancement; some compounds unexpectedly inhibited gene expression, even when blocking interferon responses.

Area of Science:

  • Biotechnology
  • Cell Biology
  • Molecular Medicine

Background:

  • In vitro transcribed (IVT) mRNA is a key tool for delivering reprogramming genes to fibroblasts for stem cell derivation.
  • Mammalian cells exhibit interferon (IFN) responses to IVT mRNA, which can reduce transfection efficiency.
  • Small molecule inhibitors of IFN have been proposed as a strategy to enhance mRNA transfection.

Purpose of the Study:

  • To screen commercially available small molecules, including known IFN inhibitors, for their ability to enhance mRNA transfection efficiency in BJ fibroblasts.
  • To evaluate the impact of these small molecules on green fluorescent protein (GFP) expression as a measure of transfection success.

Main Methods:

  • Screening of commercially available small molecules for their effect on mRNA transfection in BJ fibroblasts.
  • Quantification of transfection enhancement using relative mean fluorescence intensity of translated GFP.
  • Assessment of compound toxicity and impact on IFN-β production.

Main Results:

  • None of the tested small molecules enhanced mRNA transfection in BJ fibroblasts within toxicological limits.
  • A subset of tested compounds unexpectedly inhibited GFP expression.
  • Inhibition of IFN-β production did not correlate with enhanced GFP expression.

Conclusions:

  • Small molecule inhibitors, including those targeting IFN, do not enhance in vitro mRNA transfection efficiency in human fibroblasts.
  • The tested compounds did not overcome the cellular barriers to efficient mRNA delivery in this model.
  • Further strategies are needed to improve mRNA transfection in human fibroblasts.
Keywords:
interferonmRNA deliverysmall moleculestransfection

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