Conditional Mutation of SMC5 in Mouse Embryonic Fibroblasts

Part of the Methods in Molecular Biology book series (MIMB, volume 2004)


The structural maintenance of chromosomes (SMC) complex, SMC5/6, is important for genome maintenance in all model eukaryotes. To date, the most extensive studies have focused on the roles of Smc5/6 in lower eukaryotes, such as yeast and fly. In the handful of studies that have used mammalian cells, siRNA was used by most to knockdown SMC5/6 components. RNAi methods have been very important for scientific progression, but they are hindered by incomplete silencing of protein expression and off-target effects. This chapter outlines the use of a conditional knockout approach in mouse embryonic fibroblasts to study the function of the SMC5/6 complex. These cell lines provide an alternative method to study the function and properties of the SMC5/6 complex in mammals.

Key words

Structural maintenance of chromosomes SMC5/6 Mouse embryonic fibroblast Conditional knockout DNA damage Chromosome segregation Micronuclei DNA bridges Mitotic catastrophe 



This work was supported by the National Institutes of Health (NIH) grant K99/R00 HD069458 to P.W.J.


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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular Biology, Bloomberg School of Public HealthJohns Hopkins UniversityBaltimoreUSA

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