@article{5bd1cb4933e34f3bb0b9ffee4ed40eb8,
title = "Large-scale generation of functional mRNA-encapsulating exosomes via cellular nanoporation",
abstract = "Exosomes are attractive as nucleic-acid carriers because of their favourable pharmacokinetic and immunological properties and their ability to penetrate physiological barriers that are impermeable to synthetic drug-delivery vehicles. However, inserting exogenous nucleic acids, especially large messenger RNAs, into cell-secreted exosomes leads to low yields. Here we report a cellular-nanoporation method for the production of large quantities of exosomes containing therapeutic mRNAs and targeting peptides. We transfected various source cells with plasmid DNAs and stimulated the cells with a focal and transient electrical stimulus that promotes the release of exosomes carrying transcribed mRNAs and targeting peptides. Compared with bulk electroporation and other exosome-production strategies, cellular nanoporation produced up to 50-fold more exosomes and a more than 103-fold increase in exosomal mRNA transcripts, even from cells with low basal levels of exosome secretion. In orthotopic phosphatase and tensin homologue (PTEN)-deficient glioma mouse models, mRNA-containing exosomes restored tumour-suppressor function, enhanced inhibition of tumour growth and increased survival. Cellular nanoporation may enable the use of exosomes as a universal nucleic-acid carrier for applications requiring transcriptional manipulation.",
author = "Zhaogang Yang and Junfeng Shi and Jing Xie and Yifan Wang and Jingyao Sun and Tongzheng Liu and Yarong Zhao and Xiuting Zhao and Xinmei Wang and Yifan Ma and Veysi Malkoc and Chiling Chiang and Weiye Deng and Yuanxin Chen and Yuan Fu and Kwak, {Kwang J.} and Yamin Fan and Chen Kang and Changcheng Yin and June Rhee and Paul Bertani and Jose Otero and Wu Lu and Kyuson Yun and Lee, {Andrew S.} and Wen Jiang and Lesheng Teng and Kim, {Betty Y.S.} and Lee, {L. James}",
note = "Funding Information: This work was partially supported by the National Science Foundation of the USA (EEC-0914790, L.J.L.), the National Natural Science Foundation of China (no. 81502999, L.T. and no. 81773758, T.L.), the National Heart, Lung, and Blood Institute (R01HL132355, J.O.), the National Institute of Neurological Disorders and Stroke Grant (R01 NS104315, B.Y.S.K.), the Cancer Prevention and Research Institute of Texas (RR180017, W.J.), the American Brain Tumor Association (DG1900021) and the National Cancer Institute (K08 CA241070, W.J.). We acknowledge J. Perrino (Stanford University) for TEM imaging, which was supported in part by the National Center for Research Resources (1S10RR026780-01). This work{\textquoteright}s contents are solely the responsibility of the authors and do not necessarily represent the official views of the National Center for Research Resources or the National Institutes of Health. The authors thank D. Hollingshead at Nanotech West Lab, the Ohio State University for assisting with CNP device fabrication, X. Huang (Department of Biophysics, Peking University) for providing critical help on cryo-EM imaging, F. Meng (School of Life Sciences, Jilin University) for exosome preparation and confocal microscopy, and A. L. Chun of Science Storylab and J. Feinberg of UT Southwestern Medical Center for their editorial services.",
year = "2020",
month = jan,
day = "1",
doi = "10.1038/s41551-019-0485-1",
language = "English (US)",
volume = "4",
pages = "69--83",
journal = "Nature Biomedical Engineering",
issn = "2157-846X",
publisher = "Nature Publishing Group",
number = "1",
}