Transfection of Human Keratinocytes with Nucleoside-Modified mRNA Encoding CPD-Photolyase to Repair DNA Damage

Methods Mol Biol. 2016:1428:219-28. doi: 10.1007/978-1-4939-3625-0_14.

Abstract

In vitro-synthesized mRNA containing nucleoside modifications has great therapeutical potential to transiently express proteins with physiological importance. One such protein is photolyase which rapidly removes UV-induced DNA damages, but this enzyme is absent in humans. Here, we apply a novel mRNA-based platform to achieve functional nonhuman photolyase production in cultured human keratinocytes. Transfection of nucleoside-modified mRNA encoding photolyase leads to accelerated repair of DNA photolesions in human keratinocytes.

Keywords: Cyclobutane pyrimidine dimer-specific photolyase; DNA repair; Human keratinocytes; Nonviral gene therapy; Pseudouridine-modified mRNA; Transient transfection; Ultraviolet B.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • DNA / radiation effects
  • DNA Damage
  • DNA Repair
  • Deoxyribodipyrimidine Photo-Lyase / chemistry
  • Deoxyribodipyrimidine Photo-Lyase / genetics*
  • Deoxyribodipyrimidine Photo-Lyase / metabolism
  • Humans
  • Keratinocytes / cytology*
  • Keratinocytes / metabolism
  • Nucleosides / metabolism*
  • RNA, Messenger / chemistry*
  • RNA, Messenger / metabolism
  • Transfection

Substances

  • Nucleosides
  • RNA, Messenger
  • DNA
  • Deoxyribodipyrimidine Photo-Lyase