- Transient siRNA-mediated protein knockdown in mouse followed by feeding/starving cycle and liver tissue analysis.
Transient siRNA-mediated protein knockdown in mouse followed by feeding/starving cycle and liver tissue analysis.
STAR protocols (2021-05-18)
Lidia Wrobel, Farah H Siddiqi, David C Rubinsztein
PMID33997814
ABSTRACT
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).
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Monoclonal Anti-Tubulin, Tyrosine antibody produced in mouse, clone TUB-1A2, ascites fluid
Sigma-Aldrich
Phosphatase Inhibitor Cocktail 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)