内体
转染
荧光素酶
细胞内
信使核糖核酸
胞浆
化学
体内
核酸
细胞生物学
生物物理学
药物输送
生物
生物化学
酶
基因
生物技术
有机化学
作者
Yuhang Jiang,Qiao Lu,Yongheng Wang,Emily Xu,A. S. L. Ho,Prati Pal Singh,Yifei Wang,Zhaozhong Jiang,Fan Yang,Gregory T. Tietjen,Peter Cresswell,W. Mark Saltzman
出处
期刊:Nano Letters
[American Chemical Society]
日期:2020-01-31
卷期号:20 (2): 1117-1123
被引量:71
标识
DOI:10.1021/acs.nanolett.9b04426
摘要
Endosomal escape is a key step for intracellular drug delivery of nucleic acids, but reliable and sensitive methods for its quantitation remain an unmet need. In order to rationally optimize the mRNA transfection efficiency of a library of polymeric materials, we designed a deactivated Renilla luciferase-derived molecular probe whose activity can be restored only in the cytosol. This probe can be coencapsulated with mRNA in the same delivery vehicle, thereby accurately measuring its endosomal escape efficiency. We examined a library of poly(amine-co-ester) (PACE) polymers with different end groups using this probe and observed a strong correlation between endosomal escape and transfection efficiency (R2 = 0.9334). In addition, we found that mRNA encapsulation efficiency and endosomal escape, but not uptake, were determinant factors for transfection efficiency. The polymers with high endosomal escape/transfection efficiency in vitro also showed good transfection efficiency in vivo, and mRNA expression was primarily observed in spleens after intravenous delivery. Together, our study suggests that the luciferase probe can be used as an effective tool to quantitate endosomal escape, which is essential for rational optimization of intracellular drug delivery systems.
科研通智能强力驱动
Strongly Powered by AbleSci AI