21699593
not annotated - annotated - LINNAEUS only
Comparison of transgene expression in Aedes aegypti generated by mariner Mos1 transposition and PhiC31 site-directed recombination.
Transgenic mosquitoes generated by transposable elements (TEs) often poorly express transgenes owing to position effects. To avoid these effects, the PhiC31 site-directed recombination system was used to insert transgenes into a locus favourable for gene expression in Aedes aegypti. We describe phenotypes of mariner Mos1 TE and PhiC31 transgenic mosquitoes expressing the enhanced green fluorescent protein (EGFP) reporter in midguts of blood-fed females. Mosquitoes of nine TE-generated lines [estimated transformation frequency (TF): 9.3%] clearly expressed the eye-specific selection marker but only 2/9 lines robustly expressed the EGFP reporter. The piggyBac TE-generated PhiC31 docking strain, attP26, supported recombination with attB site containing donors at an estimated TF of 1.7-4.9%. Using a codon-optimized PhiC31 integrase mutant instead of the 'wild-type' enzyme did not affect TF. Site-directed recombination of line attP26 with an attB-containing donor expressing EGFP from the Ae. aegypti carboxypeptidase promoter produced one transgenic line with blood-fed females expressing the reporter in midgut tissue. Docking strain attP26 also supported robust expression of Flock House virus B2 from the Ae. aegypti polyubiquitin promoter. Our data confirm that eye-specific selection marker expression alone is not a reliable indicator for robust gene-of-interest expression in Ae. aegypti and that the PhiC31 system can ensure predictable transgene expression in this mosquito species.
Ann file
T1 Species 38 51 Aedes aegypti
N1 Reference T1 Taxonomy:7159
T2 Species 403 416 Aedes aegypti
N2 Reference T2 Taxonomy:7159
T3 Species 1132 1143 Ae. aegypti
N3 Reference T3 Taxonomy:7159
T4 Species 1322 1342 Flock House virus B2
N4 Reference T4 Taxonomy:12287
T5 Species 1352 1363 Ae. aegypti
N5 Reference T5 Taxonomy:7159
T6 Species 1527 1538 Ae. aegypti
N6 Reference T6 Taxonomy:7159