Figure 1. Schematic of split fluorescent protein system and insertion sites
Abstract
Visualizing the subcellular localization of presynaptic proteins with fluorescent proteins is a powerful tool to dissect the genetic and molecular mechanisms underlying synapse formation and patterning in live animals. Here, we utilize split green and red fluorescent proteins to visualize the localization of endogenously expressed presynaptic proteins at a single neuron resolution in Caenorhabditis elegans. By using CRISPR/Cas9 genome editing, we generated a collection of C. elegans strains in which endogenously expressed presynaptic proteins (RAB-3/Rab3, CLA-1/Piccolo, SYD-2/Liprin-α, UNC-10/RIM and ELKS-1/ELKS) are tagged with tandem repeats of GFP11 and/or wrmScarlet11. We show that the expression of wrmScarlet1-10 and GFP1-10 under neuron-specific promoters can robustly label presynaptic proteins in different neuron types. We believe that combinations of knock-in strains and wrmScarlet1-10 and GFP1-10 plasmids are a versatile modular system to examine the localization of endogenous presynaptic proteins in any neuron type.