Identifying chromosomal targets of DNA-binding proteins by Sequence Tag Analysis of Genomic Enrichment (STAGE)

Curr Protoc Mol Biol. 2005 Nov:Chapter 21:Unit 21.10. doi: 10.1002/0471142727.mb2110s72.

Abstract

Sequence Tag Analysis of Genomic Enrichment (STAGE) is a method for experimentally identifying the in vivo chromosomal targets of DNA-binding proteins in any sequenced genome. STAGE generates 21-bp tags derived from DNA isolated by chromatin immunoprecipitation (ChIP; UNIT 21.3). Concatamers of tags are cloned and sequenced to yield a STAGE library. Tags in the library represent DNA fragments that were occupied by the DNA-binding protein, and mapping these tag sequences to the genome identifies the binding loci of the DNA-binding protein in vivo. STAGE can be applied to any sequenced genome to identify targets of DNA-binding proteins without requiring extensive microarray resources.

MeSH terms

  • Biomedical Research / methods*
  • Chromatin Immunoprecipitation
  • Chromosome Mapping / methods*
  • Chromosomes / genetics
  • DNA-Binding Proteins / genetics*
  • Expressed Sequence Tags*
  • Genome
  • Indicators and Reagents
  • Sequence Tagged Sites*

Substances

  • DNA-Binding Proteins
  • Indicators and Reagents