Intracellular Delivery of His-Tagged Genome-Editing Proteins Enabled by Nitrilotriacetic Acid-Containing Lipidoid Nanoparticles

Adv Healthc Mater. 2019 Mar;8(6):e1800996. doi: 10.1002/adhm.201800996. Epub 2018 Nov 22.


Protein- and peptide-based therapeutics with high tolerance and specificity along with low off-target effects and genetic risks have attracted tremendous attention over the last three decades. Herein, a new type of noncationic lipidoid nanoparticle (LNP) is reported for His-tagged protein delivery. Active lipidoids are synthesized by conjugating a nitrilotriacetic acid group with hydrophobic tails (EC16, O16B, and O17O) and nanoparticles are formulated in the presence of nickel ions and helper lipids (cholesterol, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000]). It is demonstrated that the newly developed LNPs are capable of delivering various His-tagged proteins including green fluorescent protein (GFP), (-30)GFP-Cre recombinase, and CRISPR/Cas9 ribonucleoprotein into mammalian cells.

Keywords: CRISPR/Cas9; Cre recombinase; genome editing; lipidoid nanoparticles; protein delivery.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • CRISPR-Cas Systems / genetics
  • Cell Line
  • Cell Survival / drug effects
  • Gene Editing / methods*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Histidine / genetics
  • Histidine / metabolism
  • Humans
  • Lipids / chemistry*
  • Nanoparticles / chemistry*
  • Nanoparticles / toxicity
  • Nitrilotriacetic Acid / chemistry*
  • Oligopeptides / genetics
  • Oligopeptides / metabolism
  • Transfection


  • His-His-His-His-His-His
  • Lipids
  • Oligopeptides
  • Green Fluorescent Proteins
  • Histidine
  • Nitrilotriacetic Acid