Nitrilotriacetic acid–containing noncationic lipidoids are synthesized and formulated with helper lipids to form the lipidoid nanoparticles for intracellular delivery of the His‐tagged genome‐editing proteins, which take advantage of the coordination interaction between divalent nickel ion–immobilized lipidoids and the imidazole groups present on the histidine residues.
Abstract
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.
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