title: Seamless gene tagging by endonuclease-driven homologous recombination.
abstract: Gene tagging facilitates systematic genomic and proteomic analyses but chromosomal tagging typically disrupts gene regulatory sequences. Here we describe a seamless gene tagging approach that preserves endogenous gene regulation and is potentially applicable in any species with efficient DNA double-strand break repair by homologous recombination. We implement seamless tagging in Saccharomyces cerevisiae and demonstrate its application for protein tagging while preserving simultaneously upstream and downstream gene regulatory elements. Seamless tagging is compatible with high-throughput strain construction using synthetic genetic arrays (SGA), enables functional analysis of transcription antisense to open reading frames and should facilitate systematic and minimally-invasive analysis of gene functions.
authors.affiliation: Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Khmelinskii
authors.forename: Matthias
authors.initials: M
authors.lastname: Meurer
authors.forename: Nurlanbek
authors.initials: N
authors.lastname: Duishoev
authors.forename: Nicolas
authors.initials: N
authors.lastname: Delhomme
authors.forename: Michael
authors.initials: M
authors.lastname: Knop
mesh_terms: D004720:Endonucleases / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D011995:Recombination, Genetic / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
chemical_list: D004720:Endonucleases