2. Academic Validation
  3. Visualization of the process of a nanocarrier-mediated gene delivery: stabilization, endocytosis and endosomal escape of genes for intracellular spreading

Visualization of the process of a nanocarrier-mediated gene delivery: stabilization, endocytosis and endosomal escape of genes for intracellular spreading

  • J Nanobiotechnology. 2022 Mar 9;20(1):124. doi: 10.1186/s12951-022-01336-6.
Zhongzheng Ma  # 1 2,  Yang Zheng  # 3,  Zijian Chao 1,  Hongtao Chen 4,  Yunhui Zhang 1,  Meizhen Yin 4,  Jie Shen 1,  Shuo Yan 5
Affiliations

Affiliations

  • 1 Department of Plant Biosecurity and MOA Key Laboratory of Pest Monitoring and Green Management, College of Plant Protection, China Agricultural University, Beijing, 100193, People's Republic of China.
  • 2 Institute of Plant Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing, 100097, People's Republic of China.
  • 3 College of Horticulture and Plant Protection, Yangzhou University, Yangzhou, 225002, Jiangsu, People's Republic of China.
  • 4 State Key Laboratory of Chemical Resource Engineering, Beijing Lab of Biomedical Materials, Beijing University of Chemical Technology, Beijing, 100029, People's Republic of China.
  • 5 Department of Plant Biosecurity and MOA Key Laboratory of Pest Monitoring and Green Management, College of Plant Protection, China Agricultural University, Beijing, 100193, People's Republic of China. [email protected].
  • # Contributed equally.
PMID: 35264206 DOI: 10.1186/s12951-022-01336-6
Abstract

Nanoparticles have been widely applied as gene carrier for improving RNA interference (RNAi) efficiency in medical and agricultural fields. However, the mechanism and delivery process of nanoparticle-mediated RNAi is not directly visualized and elucidated. Here we synthesized a star polymer (SPc) consisted of a hydrophilic shell with positively-charged tertiary amine in the side chain, which was taken as an example to investigate the mechanism in gene delivery. The SPc could assemble with dsRNA spontaneously through electrostatic force, hydrogen bond and van der Waals force. Interestingly, the SPc could protect dsRNA from degradation by RNase A and insect hemolymph, thus remarkably increasing the stability of dsRNA. Meanwhile, the SPc could efficiently promote the cellular uptake and endosomal escape for intracellular spreading of dsRNA. Transcriptome analysis revealed that the SPc could up-regulate some key genes such as Chc, AP2S1 and Arf1 for activating clathrin-mediated endocytosis. Furthermore, the suppression of endocytosis hindered the cellular uptake of SPc-delivered dsRNA in vitro, and the subsequent RNAi effect was also disappeared in vivo. To our knowledge, our study is the first direct visualization of the detailed cellular delivery process and mechanism of nanocarrier-mediated gene delivery. Above mechanism supports the application of nanocarrier-based RNAi in gene therapy and pest management.

Keywords

Cellular uptake; Clathrin; Endocytosis; Nanocarrier; RNA interference; dsRNA.

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