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.

For plasmid and strains not available through Euroscarf, please send me an email containing an exact list of the materials.