Seamless gene tagging – Efficient site directed genome manipulations
Recently, we developed a new method for seamless gene tagging (Khmelinskii et al., 2011). This method leads to genomic insertion such that only a desired stretch of DNA (e.g. tag such as GFP), and no other foreign DNA (such as a marker gene) is introduced. This preserves all gene-adjacent regulatory sequences, such as 3’-UTR in C-terminal tagging, which contain important determinants of mRNA stability, localization or transcriptional regulation (e.g. by antisense transcription). Nevertheless, the method is fully compatible with high-throughput genetics and strain construction (e.g. using SGA technology). Currently we are using this method to construct a genome wide strain collection where each yeast gene is tagged with a fluorescent reporter in the undisturbed genomic context.
In the future we will continue to develop such and other methods; usually these undertaken emerge from an urge to manipulate cells in a specific manner where no method is available.
General resources – the pYM-cassettes for genomic PCR-targeting
We have also continuously contributed to yeast methods, where we published several (very well cited) papers that describe important new tools for yeast genomic manipulations. A methods review can be found here.
Many of our resources, in particular plasmids, are distributed by Euroscarf, a non profit organization. Please order these materials there, as we siomply cannot handle all the requests ourselves.