含氮三乙酸脂质纳米颗粒实现组蛋白标记基因组编辑蛋白的细胞内递送。
Intracellular Delivery of His-Tagged Genome-Editing Proteins Enabled by Nitrilotriacetic Acid-Containing Lipidoid Nanoparticles.
机构信息
Department of Biomedical Engineering, Tufts University, Medford, MA, 02155, USA.
出版信息
Adv Healthc Mater. 2019 Mar;8(6):e1800996. doi: 10.1002/adhm.201800996. Epub 2018 Nov 22.
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.
摘要
在过去的三十年中，具有高耐受性和特异性、低脱靶效应和遗传风险的蛋白质和肽类治疗药物引起了极大的关注。在此，报道了一种新型的非阳离子脂质纳米颗粒（LNP）用于 His 标签蛋白的递药。活性脂质体是通过将氮川三乙酸基团与疏水尾部（EC16、O16B 和 O17O）偶联合成的，纳米颗粒是在镍离子和辅助脂质（胆固醇、1,2-二油酰基-sn-甘油-3-磷酸乙醇胺和 1,2-二硬脂酰基-sn-甘油-3-磷酸乙醇胺-N-[甲氧基（聚乙二醇）-2000]）的存在下配制的。结果表明，新开发的 LNP 能够递送各种 His 标签蛋白，包括绿色荧光蛋白（GFP）、（-30）GFP-Cre 重组酶和 CRISPR/Cas9 核糖核蛋白进入哺乳动物细胞。
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