A Toolbox for Efficient Proximity-Dependent Biotinylation in Zebrafish Embryos.

ABSTRACT: Understanding how proteins are organized in compartments is essential to elucidating their function. While proximity-dependent approaches such as BioID have enabled a massive increase in information about organelles, protein complexes, and other structures in cell culture, to date there have been only a few studies on living vertebrates. Here, we adapted proximity labeling for protein discovery in vivo in the vertebrate model organism, zebrafish. Using lamin A (LMNA) as bait and green fluorescent protein (GFP) as a negative control, we developed, optimized, and benchmarked in vivo TurboID and miniTurbo labeling in early zebrafish embryos. We developed both an mRNA injection protocol and a transgenic system in which transgene expression is controlled by a heat shock promoter. In both cases,

SUBMITTER: Rosenthal SM 

PROVIDER: S-EPMC8383115 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature