Knocking in multifunctional gene tags into SMC complex subunits using gene editing

Publikation: Bidrag til bog/antologi/rapportBidrag til bog/antologiForskningfagfællebedømt

Condensin, a highly conserved pentameric chromosome complex, is required for the correct organization and folding of the genome. Here, we highlight how to knock protein tags into endogenous loci to faithfully study the condensin complex in vertebrates and dissect its multiple functions. These include using the streptavidin binding peptide (SBP) to create the first genome-wide map of condensin and perform varied applications in proteomics and enzymology of the complex. The revolution in gene editing using CRISPR/Cas9 has made it possible to insert tags into endogenous loci with relative ease, allowing physiological and fully functional tagged protein to be analyzed biochemically (affinity tags), microscopically (fluorescent tags) or both purified and localized (multifunctional tags). In this chapter, we detail how to engineer vertebrate cells using CRISPR/Cas9 to provide researchers powerful tools to obtain greater precision than ever to understand how the complex interacts and behaves in cells.

OriginalsprogEngelsk
TitelSMC Complexes : Methods and Protocols
Antal sider12
ForlagHumana Press
Publikationsdato2019
Sider91-102
ISBN (Trykt)978-1-4939-9522-6
ISBN (Elektronisk)978-1-4939-9520-2
DOI
StatusUdgivet - 2019
Eksternt udgivetJa
NavnMethods in Molecular Biology
Vol/bind2004
ISSN1064-3745

Bibliografisk note

Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2019.

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