A boronic acid-rich dendrimer with robust and unprecedented efficiency for cytosolic protein delivery and CRISPR-Cas9 gene editing

Sci Adv. 2019 Jun 12;5(6):eaaw8922. doi: 10.1126/sciadv.aaw8922. eCollection 2019 Jun.

Abstract

Cytosolic protein delivery is of central importance for the development of protein-based biotechnologies and therapeutics; however, efficient intracellular delivery of native proteins remains a challenge. Here, we reported a boronic acid-rich dendrimer with unprecedented efficiency for cytosolic delivery of native proteins. The dendrimer could bind with both negatively and positively charged proteins and efficiently delivered 13 cargo proteins into the cytosol of living cells. All the delivered proteins kept their bioactivities after cytosolic delivery. The dendrimer ensures efficient intracellular delivery of Cas9 protein into various cell lines and showed high efficiency in CRISPR-Cas9 genome editing. The rationally designed boronic acid-rich dendrimer permits the development of an efficient platform with high generality for the delivery of native proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Boronic Acids / chemistry*
  • CRISPR-Associated Protein 9 / genetics
  • CRISPR-Associated Protein 9 / metabolism
  • CRISPR-Cas Systems*
  • Cell Line, Tumor
  • Dendrimers / chemistry*
  • Dendrimers / metabolism
  • Drug Delivery Systems / methods*
  • Fluorescein-5-isothiocyanate / chemistry
  • Fluorescent Dyes / chemistry
  • Gene Editing / methods*
  • Gene Transfer Techniques*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • HeLa Cells
  • Hemoglobins / chemistry
  • Hemoglobins / genetics
  • Humans
  • Mice
  • Models, Molecular
  • NIH 3T3 Cells
  • Protein Structure, Secondary
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • RNA, Guide, CRISPR-Cas Systems / metabolism
  • Staining and Labeling / methods
  • beta Catenin / chemistry
  • beta Catenin / genetics

Substances

  • Boronic Acids
  • CTNNB1 protein, human
  • Dendrimers
  • Fluorescent Dyes
  • Hemoglobins
  • PAMAM Starburst
  • RNA, Guide, CRISPR-Cas Systems
  • beta Catenin
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • hemoglobin B
  • CRISPR-Associated Protein 9
  • Fluorescein-5-isothiocyanate