Although a plethora of gene carriers have been developed for potential gene therapy, imageable stimuli-responsive gene vectors with fast access to the nucleus, high biocompatibility, and transfection efficiency are still scarce. Herein, we report the design and synthesis of four dendrite-shaped cationic liposomes, MPA-HBI-R/DOPE (R: n-butyl, 1; n-octyl, 2; n-dodecyl, 3; palmyl, 4), prepared via esterification of 4-alkoxybenzylideneimidazolinone containing aliphatic chains of different lengths (HBI-R), the green fluorescent protein (GFP) chromophore, with a di[12]aneN₃ unit. Liposomes were fabricated via the self-assembly of MPA-HBI-R, assisted with 1,2-dioleoyl-sn-glycerol-3-phosphorylethanolamine (DOPE). These liposomes (MPA-HBI-R/DOPE) exhibited efficient DNA condensation, pH-responsive degradation, excellent cellular biocompatibility (up to 150 μM), and high transfection efficiency. Molecular docking experiments were also used to verify the optimal interaction between MPA-HBI-R and DNA, as well as the fluorescence enhancements. In particular, MPA-HBI-2/DOPE delivered DNA into the nucleus in less than an hour, and its luciferase transfection activity was more than 10 times that by Lipo2000, across multiple cell lines. The GFP chromophore conjugation allowed trackable intracellular delivery and release of DNA in real time via fluorescence imaging. Furthermore, efficient red fluorescent protein (RFP) transfection in zebrafish, with an efficiency of more than 6 times that by Lipo2000, was also achieved. The results not only realized, for the first time, the combination of gene delivery and GFP-simulated light emission, allowing fluorescent tracking and highly efficient gene transfection, but also offered valuable insights into the use of biomimetic chromophore for the development of the next-generation nonviral vectors.

中文翻译：

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Di[12]aneN3 功能化绿色荧光蛋白发色团，用于体外和体内 GFP 发光模拟和高效基因转染

尽管已经开发出大量基因载体用于潜在的基因治疗，但具有快速进入细胞核、高生物相容性和转染效率的可成像刺激响应基因载体仍然稀缺。在此，我们报告了四种树枝状阳离子脂质体MPA-HBI-R /DOPE（R：正丁基，1；正辛基，2；正十二烷基，3；棕榈基，4）的设计和合成。通过含有不同长度的脂肪链 (HBI-R) 的 4-烷氧基亚苄基咪唑啉酮、绿色荧光蛋白 (GFP) 发色团与 di[12]aneN ₃单元的酯化。脂质体被制造出来通过MPA-HBI-R的自组装，辅以 1,2-dioleoyl - sn -glycerol-3-phosphorylethanolamine (DOPE)。这些脂质体 ( MPA-HBI-R/ DOPE) 表现出高效的 DNA 浓缩、pH 响应降解、出色的细胞生物相容性（高达​​ 150 μM）和高转染效率。分子对接实验也用于验证MPA-HBI-R和 DNA 之间的最佳相互作用，以及荧光增强。特别是MPA-HBI-2/DOPE 在不到一个小时的时间内将 DNA 送入细胞核，其荧光素酶转染活性是 Lipo2000 的 10 倍以上，跨越多个细胞系。GFP 发色团结合允许通过荧光成像实时追踪细胞内 DNA 的传递和释放。此外，还实现了对斑马鱼的高效红色荧光蛋白 (RFP) 转染，其效率是 Lipo2000 的 6 倍以上。该结果不仅首次实现了基因传递和 GFP 模拟光发射的结合，实现了荧光跟踪和高效基因转染，而且还为使用仿生发色团开发下一代提供了宝贵的见解。代非病毒载体。