Effective mRNA Protection by Poly(l-ornithine) Synergizes with Endosomal Escape Functionality of a Charge-Conversion Polymer toward Maximizing mRNA Introduction Efficiency

Anjaneyulu Dirisala, Satoshi Uchida, Junjie Li, Joachim F.R. Van Guyse, Kotaro Hayashi, Sai V.C. Vummaleti, Sarandeep Kaur, Yuki Mochida, Shigeto Fukushima, Kazunori Kataoka

Research output: Contribution to journalArticlepeer-review

47 Citations (Scopus)

Abstract

For efficient delivery of messenger (m)RNA, delivery carriers need two major functions: protecting mRNA from nucleases and translocating mRNA from endolysosomes to the cytoplasm. Herein, these two complementary functionalities are integrated into a single polyplex by fine-tuning the catiomer chemical structure and incorporating the endosomal escape modality. The effect of the methylene spacer length on the catiomer side chain is evaluated by comparing poly(l-lysine) (PLL) with a tetramethylene spacer and poly(L-ornithine) (PLO) with a trimethylene spacer. Noteworthily, the nuclease stability of the mRNA/catiomer polyplexes is largely affected by the difference in one methylene group, with PLO/mRNA polyplex showing enhanced stability compared to PLL/mRNA polyplex. To introduce the endosomal escape function, the PLO/mRNA polyplex is wrapped with a charge-conversion polymer (CCP), which is negatively charged at extracellular pH but turns positive at endosomal acidic pH to disrupt the endosomal membrane. Compared to the parent PLO/mRNA polyplex, CCP facilitated the endosomal escape of the polyplex in cultured cells to improve the protein expression efficiency from mRNA by approximately 80-fold. Collectively, this system synergizes the protective effect of PLO against nucleases and the endosomal escape capability of CCP in mRNA delivery.

Original languageEnglish
Article number2100754
JournalMacromolecular rapid communications
Volume43
Issue number12
DOIs
Publication statusPublished - Jun 2022
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Materials Chemistry
  • Polymers and Plastics
  • Organic Chemistry

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