研究目的
To develop a gene delivery system using spiropyran as a photoswitchable photosensitizer for controlled photochemical internalization effect, enhancing gene delivery efficiency while minimizing photocytotoxicity.
研究成果
The study successfully developed a gene delivery system using spiropyran as a photoswitchable photosensitizer, which allows for controlled photochemical internalization effect, enhances gene delivery efficiency, and minimizes photocytotoxicity. The system demonstrates negligible dark- and photo-cytotoxicity and achieves a maximal transfection efficiency of 91%.
研究不足
The study is limited to in vitro applications, and the need for UV light to activate the system may limit its in vivo applications due to poor tissue penetration. However, the potential for in vivo application exists with near-infrared two-photon light irradiation.
1：Experimental Design and Method Selection:
The study involved the construction of ternary polyplexes by assembling spiropyran-containing polymers into the outer surface of bPEI/DNA complexes. The formation of these polyplexes was verified using fluorescence resonance energy transfer (FRET) techniques.
2：Sample Selection and Data Sources:
HeLa cells were used for in vitro transfection studies. The polyplexes were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM).
3：List of Experimental Equipment and Materials:
Equipment included Zetasizer nanoseries (Nano zs90, Malvern Instruments Ltd., U.K.), transmission electron microscopy (JEM-1011, JEOL, Ltd., Japan), and confocal laser scanning microscope (LSM 710, Zeiss, Germany). Materials included spiropyran-containing cationic copolymers PSP1 and PSP2, branched polyethylenimine (bPEI, 25 kDa), and plasmid DNA (pDNA).
4：Experimental Procedures and Operational Workflow:
The polyplexes were prepared and characterized. Intracellular ROS generation and photocytotoxicity were assessed. In vitro transfection efficiency was evaluated using flow cytometry and luciferase assays.
5：Data Analysis Methods:
Data were analyzed using FlowJo software for flow cytometry and Tanon Gis software for gel electrophoresis.
独家科研数据包，助您复现前沿成果，加速创新突破
获取完整内容
