Transient siRNA-mediated protein knockdown in mouse followed by feeding/starving cycle and liver tissue analysis.

We present a protocol for in vivo siRNA-mediated knockdown of a gene of interest in mouse liver using systemic delivery via intravenous injection. We describe a step-by-step protocol for delivery of siRNA particles, with tips on how to optimize dosage. We detail steps for feeding/starving cycles as well as for liver tissue isolation, followed by gene expression analysis, measured at the mRNA and protein levels. For complete information on the generation and use of this protocol, please refer to Wrobel et al. (2020).

Keywords

Gene Expression, Metabolism, Model Organisms, Molecular Biology, Animals, Gene Knockdown Techniques, Liver, Mice, RNA, Small Interfering, Starvation, Ubiquitin Thiolesterase

Journal Title

STAR Protoc

Journal ISSN

2666-1667
2666-1667

Volume Title

2

Publisher

Elsevier

Publisher DOI

https://doi.org/10.1016/j.xpro.2021.100500

Rights and licensing

Except where otherwised noted, this item's license is described as Attribution 4.0 International

Sponsorship

We are grateful for funding from the UK Dementia Research Institute (funded by the MRC, Alzheimer’s Research UK and the Alzheimer’s Society) (to DCR), the Roger de Spoelberch Foundation (DCR), the European Molecular Biology Organisation (EMBO long-term fellowship to LW ALTF 135-2016). We thank the University of Cambridge core mice facility staff.

Collections

University of Cambridge Research Outputs (Articles and Conferences)