Homing endonucleases catalyze double-stranded DNA breaks and somatic transgene excision in Aedes aegypti.
Aedes aegypti is a major vector of arthropod-borne viruses such as yellow fever virus and dengue viruses. Efforts to discern the function of genes involved in important behaviours, such as vector competence and host seeking through reverse genetics, would greatly benefit from the ability to generate targeted gene disruptions. Homing endonucleases are selfish elements which catalyze double-stranded DNA (dsDNA) breaks in a sequence-specific manner. In this report we demonstrate that the homing endonucleases I-PpoI, I-SceI, I-CreI and I-AniI are all able to induce dsDNA breaks in adult female Ae. aegypti chromosomes as well as catalyze the somatic excision of a transgene. These experiments provide evidence that homing endonucleases can be used to manipulate the genome of this important disease vector.
Traver
B E
BE
Department of Entomology, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Anderson
M A E
MA
Adelman
Z N
ZN
D000330:Aedes / Q000201:enzymology* / Q000235:genetics* / Q000821:virology; D000818:Animals; D001483:Base Sequence; D055162:Biocatalysis; D001681:Biological Assay; D015139:Blotting, Southern; D053903:DNA Breaks, Double-Stranded; D018983:DNA Footprinting; D004260:DNA Repair; D004274:DNA, Recombinant / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D021381:Protein Transport; D012845:Sindbis Virus / Q000502:physiology; D019076:Transgenes / Q000235:genetics*
D004274:DNA, Recombinant; D015252:Deoxyribonucleases, Type II Site-Specific