Drosophila mini-white model system: new insights into positive position effects and the role of transcriptional terminators and gypsy insulator in transgene shielding

Nucleic Acids Res. 2010 Jan;38(1):39-47. doi: 10.1093/nar/gkp877. Epub 2009 Oct 23.

Abstract

The white gene, which is responsible for eye pigmentation, is widely used to study position effects in Drosophila. As a result of insertion of P-element vectors containing mini-white without enhancers into random chromosomal sites, flies with different eye color phenotypes appear, which is usually explained by the influence of positive/negative regulatory elements located around the insertion site. We found that, in more than 70% of cases when mini-white expression was subject to positive position effects, deletion of the white promoter had no effect on eye pigmentation; in these cases, the transposon was inserted into the transcribed regions of genes. Therefore, transcription through the mini-white gene could be responsible for high levels of its expression in most of chromosomal sites. Consistently with this conclusion, transcriptional terminators proved to be efficient in protecting mini-white expression from positive position effects. On the other hand, the best characterized Drosophila gypsy insulator was poorly effective in terminating transcription and, as a consequence, only partially protected mini-white expression from these effects. Thus, to ensure maximum protection of a transgene from position effects, a perfect boundary/insulator element should combine three activities: to block enhancers, to provide a barrier between active and repressed chromatin, and to terminate transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters / genetics*
  • Animals
  • Animals, Genetically Modified
  • Chromosomal Position Effects*
  • DNA Transposable Elements
  • Drosophila / genetics*
  • Drosophila Proteins / genetics*
  • Enhancer Elements, Genetic
  • Eye Proteins / genetics*
  • Insulator Elements*
  • Models, Genetic
  • Terminator Regions, Genetic*
  • Transcription, Genetic
  • Transcriptional Activation
  • Transgenes*

Substances

  • ATP-Binding Cassette Transporters
  • DNA Transposable Elements
  • Drosophila Proteins
  • Eye Proteins
  • w protein, Drosophila
  • y protein, Drosophila