Zhang Jianxiong, Hu Yawei, Wang Xiaoqing, Liu Peng, Chen Xiaofang
Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing 100084, China.
Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China.
Micromachines (Basel). 2019 Jun 10;10(6):387. doi: 10.3390/mi10060387.
Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types.
通常需要进行细胞内基因递送以研究基因功能。需要一个能够同时进行化学转染和病毒转导以在大多数细胞类型中实现高效基因修饰的通用平台。在这里，我们证明了高通量化学转染、病毒包装和转导可以在我们之前开发的超疏水微孔阵列芯片（SMAR芯片）上高效进行。通过几个简单步骤，在芯片上物理分离的微孔中成功进行了总共169次化学转染，这有助于在SMAR芯片上进行DNA递送和更换培养基。在大幅减少人工操作的同时，实现了与传统多孔板转染相当的效率（约65%）。开发了两种转染程序，即适用于转染材料长期储存的干法和效率更高的湿法。以预定方式进行多次转染以进一步提高转染效率或在不同时间点递送多个基因。此外，还证明了与靶细胞转导相结合的高通量病毒包装，其在NIH 3T3细胞中的转基因表达效率>70%。总之，基于SMAR芯片的高通量基因递送高效且通用，可用于多种细胞类型的大规模基因修饰。
