Comparative genomics-based identification and analysis of cis-regulatory elements

Methods Mol Biol. 2012:917:245-63. doi: 10.1007/978-1-61779-992-1_15.

Abstract

Identification of cis-regulatory elements, such as enhancers and promoters, is very important not only for analysis of gene regulatory networks but also as a tool for targeted gene expression experiments. In this chapter, we introduce an easy but reliable approach to predict enhancers of a gene of interest by comparing mammalian and Xenopus genome sequences, and to examine their activity using a co-transgenesis technique in Xenopus embryos. Since the bioinformatics analysis utilizes publically available web tools, bench biologists can easily perform it without any need for special computing capability. The co-transgenesis assay, which directly uses polymerase chain reaction products, quickly screens for the activity of the candidate elements in a cloning-free manner.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Conserved Sequence
  • DNA / genetics
  • DNA / isolation & purification
  • Enhancer Elements, Genetic*
  • Gene Transfer Techniques
  • Genes, Reporter
  • Genome
  • Genomics
  • Green Fluorescent Proteins / biosynthesis
  • Green Fluorescent Proteins / genetics
  • Humans
  • Molecular Sequence Data
  • Phylogeny
  • Promoter Regions, Genetic*
  • RNA, Untranslated / genetics
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Software
  • Xenopus / genetics*
  • Xenopus Proteins / genetics

Substances

  • RNA, Untranslated
  • Xenopus Proteins
  • Green Fluorescent Proteins
  • DNA