seamless gene tagging by endonuclease-driven homologous recombination无缝的基因标记,endonuclease-driven同源重组.pdfVIP

Seamless Gene Tagging by Endonuclease-Driven Homologous Recombination 1.¤ 1.¤ 1 2 1 ¤ Anton Khmelinskii , Matthias Meurer , Nurlanbek Duishoev , Nicolas Delhomme , Michael Knop * 1 Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany, 2 Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany 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. Citation: Khmelinskii A, Meurer M, Duishoev N, Delhomme N, Knop M (2011) Seamless Gene Tagging by Endonuclease-Driven Homologous Recombination. PLoS ONE 6(8): e23794. doi:10.1371/journal.pone.0023794 Editor: Kevin G. Hardwick, University of Edinburgh, United Kingdom Received June 24, 2011; Accepted July 27, 2011; Published August 22, 2011 Copyright: 2011 Khmelinskii et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits