Targeting endogenous proteins for spatial and temporal knockdown using auxin-inducible degron in Caenorhabditis elegans
Mizuki Kurashina, Kota Mizumoto
Abstract
The auxin-inducible degron (AID) provides reversible, spatiotemporal control for the knockdown of target proteins. Here, we present a protocol for AID-mediated protein knockdown in Caenorhabditis elegans. We describe steps for generating the knock-in mutants using two CRISPR-Cas9 genome editing techniques and preparing the auxin-containing nematode growth media (NGM) plates. We also detail AID-mediated spatiotemporal protein knockdown. For complete details on the use and execution of this protocol, please refer to Kurashina et al. (2021).1
Topics & Concepts
DegronCaenorhabditis elegansGene knockdownEndogenyCell biologyBiologyGeneticsGeneBiochemistryUbiquitin ligaseUbiquitinGenetics, Aging, and Longevity in Model OrganismsCRISPR and Genetic EngineeringNeurobiology and Insect Physiology Research