Specific developmental gene silencing in the honey bee using a homeobox motif.
Manipulating the expression of genes in species that are not currently used as genetic models will provide comparative insights into the evolution of gene functions. However the experimental tools in doing so are limited in species that have not served as models for genetic studies. We have examined the effects of double stranded RNA (dsRNA) in the honey bee, an insect with considerably basic scientific interest. dsRNA derived from a 300 bp stretch of the E30 homeobox motif was injected into honey bee embryos at the anterior pole in the preblastoderm stage. We found that the dsRNA fragment successfully disrupted the protein expression of the target gene throughout the whole embryo. The disruption caused deficient phenotypes similar to known loss of function mutants of Drosophila engrailed, whereas embryos injected with nonsense dsRNA showed no abnormalities. We show that the large size of the honey bee egg (D: 0.3 mm, L: 1.6 mm) and the long preblastoderm stage (11-12 h) can be exploited to generate embryos with partial disruption of gene function, which may provide an elegant alternative to classical chimeric analyses. This is the first report of targeted disruption of gene function in the honey bee, and the results prove that the chosen target gene is a functional ortholog to engrailed in Drosophila.
Beye
M
M
Martin-Luther-Universität Halle/Wittenberg, Biozentrum, Institut für Zoologie, Molekulare Okologie, Halle, Germany. beye@zoologie.uni-halle.de
Härtel
S
S
Hagen
A
A
Hasselmann
M
M
Omholt
S W
SW
D000818:Animals; D017123:Bacteriophage T7 / Q000235:genetics; D001483:Base Sequence; D001516:Bees / Q000196:embryology / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000502:physiology; D005786:Gene Expression Regulation; D020868:Gene Silencing; D005801:Genes, Homeobox; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic