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A modular system to label endogenous presynaptic proteins using split fluorophores in C. elegans. | LitMetric

A modular system to label endogenous presynaptic proteins using split fluorophores in C. elegans.

Genetics

Graduate Program of Cell and Developmental Biology, Life Sciences Institute, The University of British Columbia, Vancouver, Canada, V6T 1Z3.

Published: December 2024

AI Article Synopsis

Article 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, SNG-1/Synaptogyrin, CLA-1/Piccolo, SYD-2/Liprin-α, UNC-10/RIM, RIMB-1/RIM-BP, and ELKS-1/ELKS) are tagged with tandem repeats of GFP11 and/or wrmScarlet11. We show that the expression of GFP1-10 and wrmScarlet1-10 under neuron-specific promoters can robustly label presynaptic proteins in different neuron types. We believe that the combination of our knock-in strains and GFP1-10 and wrmScarlet1-10 plasmids is a versatile modular system useful for neuroscientists to examine the localization of endogenous presynaptic proteins in any neuron type in C. elegans.

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Source
http://dx.doi.org/10.1093/genetics/iyae214DOI Listing

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