Modern Tools to Study Nuclear Pore Complexes and Nucleocytoplasmic Transport in Caenorhabditis elegans

• Published in: Methods in Cell Biology Vol. 122; pp. 277 - 310
• Main Authors: Askjaer, Peter, Galy, Vincent, Meister, Peter
• Format: Book Chapter Journal Article
• Language: English
• Published: United States Elsevier Science & Technology 2014; ELSEVIER ACADEMIC PRESS INC
• Series: Methods in Cell Biology
• Subjects: Active Transport, Cell Nucleus; Animals; Caenorhabditis elegans; Caenorhabditis elegans - genetics; Caenorhabditis elegans Proteins - metabolism; Cellular biology; Chromatin - metabolism; CRISPR-Cas9; Development Biology; Embryo, Nonmammalian - cytology; Embryo, Nonmammalian - embryology; Fluorescent Antibody Technique - methods; Life Sciences; Live imaging; Microscopy, Electron, Transmission - methods; npc; npp; Nuclear Pore - metabolism; Nuclear pore complex; Nuclear Pore Complex Proteins - analysis; Nucleocytoplasmic transport; Nucleoporin; RNA Interference; RNA, Small Interfering; RNAi; Super-resolution microscopy; npc; Super-resolution microscopy; Caenorhabditis elegans; RNAi; Nuclear pore complex; CRISPR-Cas9; Live imaging; Nucleocytoplasmic transport; Nucleoporin; npp
• ISBN: 9780124171602; 0124171605
• ISSN: 0091-679X
• DOI: 10.1016/B978-0-12-417160-2.00013-8

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.