TY  - GEN
T1  - Modified auxin improves the auxin-inducible degradation (AID) system for laid C. elegans embryos
AU  - Negishi, Takefumi
AU  - Asakawa, Masayo
AU  - Kanemaki, Masato T
AU  - Sawa, Hitoshi
DO  - 10.17912/micropub.biology.000190
UR  - http://beta.micropublication.org/journals/biology/micropub-biology-000190/
AB  - The targeted protein degradation systems in which a protein accompanying with specific tags can be degraded are developed as an approach of conditional loss of function analyses (Natsume and Kanemaki 2017). The insertion of tags into the gene loci by the CRISPR/Cas9 system allows us to deplete endogenous proteins in stage and cell specific manners. So far, four systems can be used to deplete the tagged proteins in C . elegans (Armenti et al. 2014; Zhang et al. 2015; Wang et al. 2017; Wu et al. 2017). The auxin-inducible degradation (AID) system can degrade the tagged protein by administration of the phytohormone auxin. In the AID system, a plant-derived TIR1 F-box protein can form an E3 ubiquitin ligase complex with the endogenous Skp and Cullin proteins, and in the presence of auxin, the complex interacts with a degron tag derived from the IAA17 transcriptional repressor (Nishimura et al. 2009; Yesbolatova et al. 2019). Consequently, the degron-tagged proteins are polyubiquitinated for degradation by the proteasome. With the controlled expression of TIR1 and administration of auxin, the AID system can allow us to perform spaciotemporal protein depletion. Although this system can be a powerful tool for conditional loss of function analyses, it appears to be difficult to degrade proteins in embryos, especially at late embryonic stages, since efficiencies of degradation in laid embryos surrounded by the eggshell is low compared to those in hatched larvae or adults (Zhang et al. 2015).
PY  - 2019
JO  - microPublication Biology
ER  -