Exogenous gypsy insulator sequences modulate transgene expression in the malaria vector mosquito, Anopheles stephensi

Proc Natl Acad Sci U S A. 2013 Apr 30;110(18):7176-81. doi: 10.1073/pnas.1304722110. Epub 2013 Apr 12.

Abstract

Malaria parasites are transmitted to humans by mosquitoes of the genus Anopheles, and these insects are the targets of innovative vector control programs. Proposed approaches include the use of genetic strategies based on transgenic mosquitoes to suppress or modify vector populations. Although substantial advances have been made in engineering resistant mosquito strains, limited efforts have been made in refining mosquito transgene expression, in particular attenuating the effects of insertions sites, which can result in variations in phenotypes and impacts on fitness due to the random integration of transposon constructs. A promising strategy to mitigate position effects is the identification of insulator or boundary DNA elements that could be used to isolate transgenes from the effects of their genomic environment. We applied quantitative approaches that show that exogenous insulator-like DNA derived from the Drosophila melanogaster gypsy retrotransposon can increase and stabilize transgene expression in transposon-mediated random insertions and recombinase-catalyzed, site-specific integrations in the malaria vector mosquito, Anopheles stephensi. These sequences can contribute to precise expression of transgenes in mosquitoes engineered for both basic and applied goals.

Publication types

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

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Anopheles / genetics*
  • Base Sequence
  • DNA / genetics
  • Drosophila melanogaster / genetics
  • Gene Expression*
  • Genes, Insect / genetics
  • Genetic Markers
  • Green Fluorescent Proteins / metabolism
  • Insect Proteins / genetics
  • Insect Proteins / metabolism
  • Insect Vectors / genetics*
  • Insulator Elements / genetics*
  • Luminescent Proteins / metabolism
  • Malaria / parasitology*
  • Molecular Sequence Data
  • Mutagenesis, Insertional / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Transgenes / genetics*
  • Transposases / metabolism

Substances

  • Cyan Fluorescent Protein
  • Genetic Markers
  • Insect Proteins
  • Luminescent Proteins
  • RNA, Messenger
  • fluorescent protein 583
  • Green Fluorescent Proteins
  • DNA
  • Transposases

Associated data

  • GENBANK/KC733875