Conditional Mutation of SMC5 in Mouse Embryonic Fibroblasts
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 wordsStructural 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.
- 16.Herbert AD, Carr AM, Hoffmann E (2014) FindFoci: a focus detection algorithm with automated parameter training that closely matches human assignments, reduces human inconsistencies and increases speed of analysis. PLoS One 9:e114749. https://doi.org/10.1371/journal.pone.0114749 CrossRefPubMedPubMedCentralGoogle Scholar