Intracranial delivery of antisense locked nucleic acids for silencing microRNAs in the mouse brain

Nikhil Sharma; Ganes C Sen

doi:10.1016/j.xpro.2022.101836

Intracranial delivery of antisense locked nucleic acids for silencing microRNAs in the mouse brain

STAR Protoc. 2022 Dec 16;3(4):101836. doi: 10.1016/j.xpro.2022.101836. Epub 2022 Nov 18.

Authors

Nikhil Sharma¹, Ganes C Sen²

Affiliations

¹ Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
² Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, 9500 Euclid Avenue, Cleveland, OH 44195, USA. Electronic address: seng@ccf.org.

Abstract

Antisense locked nucleic acid (LNA) technology has been widely used for silencing microRNAs with enhanced specificity and efficiency. In this protocol, we first describe the procedure for targeted intracranial delivery of LNAs to silence microRNAs specifically in the mouse brain. We then detail the steps to isolate RNA and protein from mouse brain, followed by using RT-PCR and Western blotting to confirm microRNA silencing. This noninvasive approach can only be applied to mouse brain to specifically target silencing of microRNAs. For complete details on the use and execution of this protocol, please refer to Sharma et al. (2021).¹.

Keywords: Gene expression; Model organisms; Molecular biology; Neuroscience.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Blotting, Western
Brain
Mice
MicroRNAs* / genetics
Oligonucleotides, Antisense / genetics

Substances

MicroRNAs
Oligonucleotides, Antisense