Key Laboratory of Radiopharmaceutics, Ministry of Education, College of Chemistry, Beijing Normal University, Beijing 100875, China.
Lab for Bone Metabolism, Key Lab for Space Biosciences and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, Shaanxi 710072, China.
ACS Appl Mater Interfaces. 2020 Sep 9;12(36):40094-40107. doi: 10.1021/acsami.0c10718. Epub 2020 Aug 26.
Although a plethora of nonviral gene vectors have been developed for potential gene therapy, imageable gemini surfactants with stimuli-responsiveness and high transfection efficiency are still scarce for gene delivery. Herein, three gemini amphiphiles () consisting of an aggregation-induced emission (AIE) central fluorophore: 5,6-diphenylpyrazine-2,3-diester (), decorated with triazole-[12]aneN as the hydrophilic moiety and alkyl chains of various lengths as the hydrophobic moiety, were designed and synthesized for trackable gene delivery via optical imaging. All three amphiphiles exhibited ultralow critical micelle concentrations (CMCs) (up to 3.40 × 10 M), prominent two-photon absorption properties, and solvatochromic fluorescence. Gel electrophoresis assays demonstrated that the migration of plasmid DNA was completely retarded after condensation with these gemini amphiphiles at low concentrations (up to 10 μM). In addition, the ester bond in these amphiphiles may facilitate vector degradation and DNA release, in response to esterase and the acidic environment inside cells. Upon self-assembly with DOPE to form liposomes, DOPE achieved the best transfection efficiency in four cell lines, and the transfection efficiency of DOPE in HeLa cell lines was 23.5-fold higher than that of Lipo2000, which is unusually high for small organic molecule-based nonviral vectors. Furthermore, excellent transfection efficiency of DOPE was obtained in the presence of serum, and the red fluorescence protein (RFP) gene was successfully transfected in zebrafish embryos. Both one- and two-photon fluorescence imaging clearly demonstrated the delivery process of plasmid DNA. This study demonstrated that gemini-type amphiphiles composed of a two-photon fluorophore core conjugated with triazole-[12]aneN via an ester bond afforded an unprecedentedly high transfection efficiency with excellent biocompatibility, which may provide new insights for the design and development of multifunctional nonviral gene vectors for imageable gene delivery.
尽管已经开发出了许多非病毒基因载体用于潜在的基因治疗，但具有刺激响应性和高转染效率的可成像双子表面活性剂仍然稀缺，难以用于基因递送。在此，设计并合成了三种双子两亲物（），由聚集诱导发射（AIE）的中央荧光团：5,6-二苯基吡嗪-2,3-二酯（）组成，用三唑-[12]烷 N 作为亲水部分，用各种长度的烷基链作为疏水部分进行修饰，用于通过光学成像进行可追踪的基因递送。所有三种两亲物都表现出超低临界胶束浓度（CMC）（高达 3.40×10 M）、显著的双光子吸收特性和溶剂致变色荧光。凝胶电泳实验表明，这些双子两亲物在低浓度（高达 10 μM）下与质粒 DNA 缩合后，完全阻止了质粒 DNA 的迁移。此外，这些两亲物中的酯键可能有利于载体的降解和 DNA 的释放，以响应酯酶和细胞内的酸性环境。与 DOPE 自组装形成脂质体后，DOPE 在四种细胞系中达到最佳转染效率，而 DOPE 在 HeLa 细胞系中的转染效率比 Lipo2000 高 23.5 倍，这对于基于小分子的非病毒载体来说是异常高的。此外，在存在血清的情况下，DOPE 也获得了优异的转染效率，并且成功地将红色荧光蛋白（RFP）基因转染到斑马鱼胚胎中。单光子和双光子荧光成像清楚地证明了质粒 DNA 的递药过程。本研究表明，由双光子荧光团核心通过酯键与三唑-[12]烷 N 连接的双子型两亲物提供了前所未有的高转染效率和优异的生物相容性，这可能为设计和开发用于可成像基因递送的多功能非病毒基因载体提供新的思路。
