Abstract
Efficient delivery of genetic material to primary cells remains challenging. Here, efficient transfer of genetic material is presented using synthetic biodegradable nanocarriers, resembling extracellular vesicles in their biomechanical properties. This is based on two main technological achievements: generation of soft biodegradable polyelectrolyte capsules in nanosize and efficient application of the nanocapsules for co-transfer of different RNAs to tumor cell lines and primary cells, including hematopoietic progenitor cells and primary T cells. Near to 100% efficiency is reached using only 2.5 × 10-4 pmol of siRNA, and 1 × 10-3 nmol of mRNA per cell, which is several magnitude orders below the amounts reported for any of methods published so far. The data show that biodegradable nanocapsules represent a universal and highly efficient biomimetic platform for the transfer of genetic material with the utmost potential to revolutionize gene transfer technology in vitro and in vivo.
Highlights
Efficient delivery of genetic material to primary cells remains challenging
Quantitative analysis based on calculation of the intensity of the green fluorescence signal revealed 80% reduction of green fluorescence in the cells treated with nanocapsules filled with GFPsiRNA and only 21% reduction of GFP signal in the cells transfected with Lipofectamine 2000; no unspecific effects were observed by the application of capsules loaded with the control siRNA (Figure S3, Supporting Information)
These results suggest that biodegradable nanocapsules possess utmost high transfer efficiency of RNA molecules with no toxic effect
Summary
Efficient delivery of genetic material to primary cells remains challenging. Here, efficient transfer of genetic material is presented using synthetic biodegradable nanocarriers, resembling extracellular vesicles in their biomechanical properties. Quantitative analysis based on calculation of the intensity of the green fluorescence signal revealed 80% reduction of green fluorescence in the cells treated with nanocapsules filled with GFPsiRNA and only 21% reduction of GFP signal in the cells transfected with Lipofectamine 2000; no unspecific effects were observed by the application of capsules loaded with the control siRNA (Figure S3, Supporting Information).
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