Modern tools to study nuclear pore complexes and nucleocytoplasmic transport in Caenorhabditis elegans

Abstract

The nematode Caenorhabditis elegans is characterized by many features that make it highly attractive to study nuclear pore complexes (NPCs) and nucleocytoplasmic transport. NPC composition and structure are highly conserved in nematodes and being amenable to a variety of genetic manipulations, key aspects of nuclear envelope dynamics can be observed in great details during breakdown, reassembly, and interphase. In this chapter, we provide an overview of some of the most relevant modern techniques that allow researchers unfamiliar with C. elegans to embark on studies of nucleoporins in an intact organism through its development from zygote to aging adult. We focus on methods relevant to generate loss-of-function phenotypes and their analysis by advanced microscopy. Extensive references to available reagents, such as mutants, transgenic strains, and antibodies are equally useful to scientists with or without prior C. elegans or nucleoporin experience.

Keywords: CRISPR-Cas9; Caenorhabditis elegans; Live imaging; Nuclear pore complex; Nucleocytoplasmic transport; Nucleoporin; RNAi; Super-resolution microscopy; npc; npp.