Single-Plasmid-Based System for Efficient Noncanonical Amino Acid Mutagenesis in Cultured Mammalian Cells

Sarit Cohen, Eyal Arbely

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

We describe a new expression system for efficient non-canonical amino acid mutagenesis in cultured mammalian cells by using the pyrrolysine tRNA synthetase/tRNACUAPyl pair. A significant improvement in the incorporation of non-canonical amino acids into proteins was obtained by combining all the required genetic components into a single and compact vector that can be efficiently delivered to different mammalian cell lines by conventional transfection reagents. Less is more: In cultured mammalian cells, the efficiency of non-canonical amino acid mutagenesis by amber suppression is usually low. We achieved up to 20-fold improvement in expression by combining all the required genetic material into single and compact expression vector.

Original languageEnglish
Pages (from-to)1008-1011
Number of pages4
JournalChemBioChem
Volume17
Issue number11
DOIs
StatePublished - 2 Jun 2016

Keywords

  • amber suppression
  • genetic code expansion
  • protein engineering
  • protein expression
  • pyrrolysine

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