Effective delivery with enhanced translational activity synergistically accelerates mRNA-based transfection

F. T. Zohra, E. H. Chowdhury, S. Tada, T. Hoshiba, T. Akaike

Research output: Contribution to journalArticleResearchpeer-review

42 Citations (Scopus)

Abstract

mRNA instead of DNA provides a new and attractive approach for gene therapy and genetic vaccination. Current technologies for mRNA delivery are based on cationic lipids with DOTAP being the most efficient one. We previously reported on the synthesis of an inorganic-organic hybrid carrier by embedding inorganic nano-particles of carbonate apatite onto liposomal carrier DOTAP and demonstrated its high transfection potency of luciferase mRNA both in mitotic and non-mitotic cells. Here we show that in addition to the carrier design for effective endocytosis and release of mRNA to the cytoplasm, enhancement of mRNA translation efficiency is a prerequisite for maximum protein expression. We used the modified cap analog (ARCA) during in vitro transcription of luciferase DNA for proper cap orientation and demonstrated that transfection with ARCA-mRNA resulted in higher protein expression than the mRNA with usual cap structure for both DOTAP and DOTAP-apatite complex. Secondly, exogenous poly(A) was co-delivered with mRNA by the DOTAP-apatite, resulting in very significant expression compared to mRNA delivery only. Finally, when combined both of the effects of smart carrier and the modifications at mRNA translational level, a notable enhancement (100 times) was achieved as compared to the existing DOTAP-based liposome technology. Our findings, therefore, unveiled a novel approach that an effective delivery system can be developed by the improvement of the gene expression level in combination with the enhancement of the carrier potency.

Original languageEnglish
Pages (from-to)373-378
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume358
Issue number1
DOIs
Publication statusPublished - 22 Jun 2007
Externally publishedYes

Keywords

  • Anti reverse cap analog
  • Carbonate apatite
  • Exogenous poly(A)
  • Inorganic-organic hybrid carrier
  • Liposome
  • mRNA delivery
  • Transgene expression

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