Drosophila mini-white model system: new insights into positive position effects and the role of transcriptional terminators and gypsy insulator in transgene shielding.
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.
Silicheva
Margarita
M
Department of the Control of Genetic Processes, Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
Golovnin
Anton
A
Pomerantseva
Ekaterina
E
Parshikov
Aleksander
A
Georgiev
Pavel
P
Maksimenko
Oksana
O
D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D055012:Chromosomal Position Effects; D004251:DNA Transposable Elements; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005136:Eye Proteins / Q000235:genetics*; D038101:Insulator Elements; D008957:Models, Genetic; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D019076:Transgenes
D018528:ATP-Binding Cassette Transporters; D004251:DNA Transposable Elements; D029721:Drosophila Proteins; D005136:Eye Proteins; C059254:w protein, Drosophila; C050680:y protein, Drosophila