Protocol for In Vivo Evaluation and Use of Destabilizing Domains in the Eye, Liver, and Beyond

STAR Protoc. 2020 Sep 18;1(2):100094. doi: 10.1016/j.xpro.2020.100094. Epub 2020 Aug 27.

Abstract

Destabilizing domains (DDs) have been used successfully to conditionally control the abundance of proteins of interest (POIs) in a small-molecule-dependent manner in mice, worms (Caenorhabditis elegans), and Drosophila. However, development of such systems must account for delivery of the DD-POIs to the target tissue, accessibility of the target tissue to the small molecule, and quantification of stabilization. Here, we describe the considerations and steps to take in order to effectively implement a DD-POI in mouse ocular and hepatic tissue. For complete details on the use and execution of this protocol, please refer to Datta et al. (2018), Ramadurgum and Hulleman (2020), and Ramadurgum et al. (2020).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Drug Delivery Systems / methods*
  • Eye / metabolism
  • Intravitreal Injections / methods
  • Liver / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Protein Domains / genetics
  • Protein Domains / physiology
  • Protein Stability / drug effects*
  • Proteolysis / drug effects*