Dmg peg 2000

DOPE and 1,2-dimyristoyl-rac-glycero-3-methylpolyoxyethylene 2000 (DMG-PEG 2000) were purchased from the NOF Corporation (Tokyo, Japan). SM derived from porcine brain was purchased from Avanti Polar Lipids (Alabaster, AL, USA). Ubiquinone-10 (CoQ₁₀) was purchased from FUJIFILM Wako Pure Chemical Co (Osaka, Japan). Stearyl-R8 was purchased from Toray Industries, Inc (Tokyo, Japan). DiD was purchased from Funakoshi Co., Ltd (Tokyo, Japan). PBS (−) tablets were purchased from Takara Bio Inc (Shiga, Japan).

pCpG-free-TgGRA15 encapsulation with LNPs was prepared by the ethanol dilution method, as defined by Maeta et al. [17 (link)], leading to the generation of ssPalmE-LNP_TgGRA15. Briefly, 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) was purchased from Avanti Polar Lipids (Alabaster, AL, USA). Cholesterol was purchased from Sigma Aldrich (St. Louis, MO, USA). The 1-(Monomethoxy poly (ethylene glycol)2000)-2,3-dimyristoylglycerol (DMG-PEG2000) and ssPalm with vitamin E as a hydrophobic scaffold (ssPalmE: COATSOME^® SS-E) were purchased from NOF corporation (Tokyo, Japan). Protamine sulfate salmon milt was purchased from Calbiochem (Darmstadt, Germany). The LNP encapsulating DNA were prepared by formation of pDNA/protamine complex, followed by encapsulation in the LNP (ssPalmE/DOPE/Chol/DMG-PEG2000 = 3/4/3/0.3). The size of the ssPalmE-LNP particle was 140 nm, and slightly negatively charged (approximately −5 mV). Moreover, the encapsulation efficiency was >90%.