Using translational enhancers to increase transgene expression in Drosophila

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 109; no. 17; pp. 6626 - 6631
• Main Authors: Pfeiffer, Barret D, Truman, James W, Rubin, Gerald M
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 24.04.2012; National Acad Sciences
• Subjects: 3' Untranslated Regions; 5' Untranslated Regions; Animals; Base Sequence; Biological Sciences; Drosophila; Drosophila - genetics; Enhancer Elements, Genetic; Enhancers; Fluorescence; Gene expression; Gene Expression Regulation; Genes; Genetics; Genomes; Genotypes; Germ cells; half life; image analysis; Inactivators; insect viruses; Insects; Messenger RNA; Molecular Sequence Data; mRNA; mRNA stability; Nervous system; Neurons; Plant viruses; Protein Biosynthesis; Proteins; RNA transport; RNA, Messenger - genetics; RNA-protein interactions; Transcription; Transgenes; Transgenic animals; Translation; translation (genetics); Untranslated regions
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1204520109

The ability to specify the expression levels of exogenous genes inserted in the genomes of transgenic animals is critical for the success of a wide variety of experimental manipulations. Protein production can be regulated at the level of transcription, mRNA transport, mRNA half-life, or translation efficiency. In this report, we show that several well-characterized sequence elements derived from plant and insect viruses are able to function in Drosophila to increase the apparent translational efficiency of mRNAs by as much as 20-fold. These increases render expression levels sufficient for genetic constructs previously requiring multiple copies to be effective in single copy, including constructs expressing the temperature-sensitive inactivator of neuronal function Shibirets1, and for the use of cytoplasmic GFP to image the fine processes of neurons.