Stimulating Macropinocytosis for Intracellular Nucleic Acid and Protein Delivery: A Combined Strategy with Membrane-Lytic Peptides To Facilitate Endosomal Escape

Bioconjug Chem. 2020 Mar 18;31(3):547-553. doi: 10.1021/acs.bioconjchem.0c00064. Epub 2020 Feb 7.

Abstract

Delivery of biomacromolecules via endocytic pathways requires the efficient accumulation of cargo molecules into endosomes, followed by their release to the cytosol. We propose a unique intracellular delivery strategy for bioactive molecules using a new potent macropinocytosis-inducing peptide derived from stromal-derived factor 1α (SN21). This peptide allowed extracellular materials to enter cells through the activation of macropinocytosis. To provide the ability to release internalized cargoes from endosomes, we conjugated SN21 with membrane-lytic peptides. The combination of a macropinocytosis-inducing peptide and a membrane-lytic peptide successfully delivered functional siRNA and proteins, which include antibodies, Cre recombinase, and an artificial transcription regulator protein having a transcription activator-like effector (TALE) motif. This study shows the feasibility of combining the physiological stimulation of macropinocytosis with the physicochemical disruption of endosomes as a strategy for intracellular delivery.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Membrane / drug effects*
  • Cell Membrane / metabolism
  • Endosomes / drug effects*
  • Endosomes / metabolism*
  • HeLa Cells
  • Humans
  • Peptides / pharmacology*
  • Pinocytosis / drug effects*
  • Protein Transport / drug effects
  • Proteins / chemistry
  • Proteins / metabolism*
  • RNA, Small Interfering / metabolism*

Substances

  • Peptides
  • Proteins
  • RNA, Small Interfering