Evaluation of microRNA delivery in vivo

Rikki A.M. Brown, Kirsty L. Richardson, Felicity C. Kalinowski, Michael R. Epis, Jessica L. Horsham, Tasnuva D. Kabir, Marisa H. De Pinho, Dianne J. Beveridge, Lisa M. Stuart, Larissa C. Wintle, Peter J. Leedman

Research output: Chapter in Book/Conference paperChapter

6 Citations (Scopus)

Abstract

MicroRNAs (miRNAs) are a family of short noncoding RNA molecules that fine-tune expression of mRNAs. Often their altered expression is associated with a number of diseases, including cancer. Given that miRNAs target multiple genes and “difficult to drug” oncogenes, they present attractive candidates to manipulate as an anti-cancer strategy. MicroRNA-7 (miR-7) is a tumor suppressor miRNA that has been shown to target oncogenes overexpressed in cancers, such as the epidermal growth factor receptor (EGFR) and the nuclear factor-κ B subunit, RelA. Here, we describe methods for evaluating systemic delivery of miR-7 using a lipid nanoparticle formulation in an animal model. The microRNA is delivered three times, over 1 week and tissues collected 24 h after the last injection. RNA and protein are extracted from snap frozen tissues and processed to detect miRNA distribution and subsequent assessment of downstream targets and signaling mediators, respectively. Importantly, variability in efficiency of miRNA delivery will be observed between organs of the same animal and also between animals. Additionally, delivering the microRNA to organs other than the liver, particularly the brain, remains challenging. Furthermore, large variation in miRNA targets is seen both within tissues and across tissues depending on the lysis buffer used for protein extraction. Therefore, analyzing protein expression is dependent upon the method used for isolation and requires optimization for each individual application. Together, these methods will provide a foundation for those planning on assessing the efficacy of delivery of a miRNA in vivo.

Original languageEnglish
Title of host publicationMicroRNA and Cancer
Subtitle of host publicationMethods and Protocols
EditorsWei Wu
PublisherHumana Press
Pages155-178
Number of pages24
Volume1699
Edition2nd
ISBN (Electronic) 9781493974351
ISBN (Print)9781493974337
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1699
ISSN (Print)1064-3745

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Brown, R. A. M., Richardson, K. L., Kalinowski, F. C., Epis, M. R., Horsham, J. L., Kabir, T. D., ... Leedman, P. J. (2018). Evaluation of microRNA delivery in vivo. In W. Wu (Ed.), MicroRNA and Cancer: Methods and Protocols (2nd ed., Vol. 1699, pp. 155-178). (Methods in Molecular Biology; Vol. 1699). Humana Press. https://doi.org/10.1007/978-1-4939-7435-1_12