Use of protein biotinylation in vivo for chromatin immunoprecipitation.
We describe a system designed to express biotinylated proteins in mammalian cells in vivo and its application to the study of protein-DNA interactions in vivo by chromatin immunoprecipitation (ChIP). The system is based on coexpression of the target protein fused to a short biotin acceptor domain together with the biotinylating enzyme BirA from Escherichia coli. The superior strength of the biotin-avidin interaction allows one to employ more stringent washing conditions in the ChIP protocol, resulting in a better signal/noise ratio.
Viens
Antoine
A
Laboratoire Oncogénse, Différenciation et Transduction du Signal, Institut André Lwoff, 7 rue Guy Moquet, 94800 Villejuif, France.
Mechold
Undine
U
Lehrmann
Heike
H
Harel-Bellan
Annick
A
Ogryzko
Vasily
V
D000818:Animals; D019921:Biotinylation; D019731:Carbon-Nitrogen Ligases / Q000737:chemistry* / Q000235:genetics; D018797:Cell Cycle Proteins / Q000235:genetics; D002843:Chromatin / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D007164:Immunosorbents; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics; D050680:Transcription Factor DP1; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics; D014161:Transduction, Genetic
D018797:Cell Cycle Proteins; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D007164:Immunosorbents; D008164:Luminescent Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D050680:Transcription Factor DP1; D014157:Transcription Factors; D000090266:Chromobox Protein Homolog 5; D049452:Green Fluorescent Proteins; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli