ALL STERILE MALES OF CULICINE MOSQUITOES: A METHOD OF CREATION

摘要

An all male Culicide mosquito population is created by knocking down its Transformer-2 gene, causing the dysfunction of X chromosome-bearing sperm, hence producing severe biased male progenies. Unlike previous methods, we recently discovered that the Tra-2 knockdown also results in female-specific zygotes lethality (XX). This art is therefore also designed to kill early female zygotes (XX) that may have survived the previous knockdown, and the all male progenies are created only when an antibiotic substance has been added into food and drink to feed mosquitoes. The strict limit of the antibiotic exposure time allows mosquito-adapted Wolbachia bacteria to survive. Selected Wolbachia bacteria may induce cytoplasmic incompatibility (CI) of up to 100%. All the progenies are therefore genetically males, which cause sterility when outcrossing with females infected by another Wolbachia strain (bidirectional CI) or are uninfected (unidirectional CI) in natural environment.

主权项

1. A method of creation of an all-male sterile Culicine mosquito population therefore, comprising:
i) creating a biological strain of Culicinae mosquito, the strain of mosquito carrying one or more insertion events of a Tet-on+Transformer-2 genetic knockdown system to silence the endogenous Tra-2 gene, wherein this knockdown effect occurring only in the said rearing conditions (laboratory or insectary), the knockdown system only being activated via the additive substance and the Tra-2 knockdown system being expressed during male spermatogenesis and killing X(m) chromosome bearing sperms and being expressed to kill early female zygotes when some X(m) chromosome-bearing sperms survived and all types of the knockdown effects producing a severe male bias, up to 100% male in the progeny; ii) the said breeding strain being under the absence of a substance, the genetic knockdown systems therefore being silent in the natural habitat of the species, the substance being absent in the natural habitat and present only when being fed to the transformant insects and inducing the knockdown system; iii) the insect strain also carrying one or more endo-symbiotic Wolbachia, wherein female infected insects being crossed with males of a Tet-on+Tra-2 RNAi strain, or embryonic cytoplasm of infected insect, is microinjected into adult female thoraxic muscles of target species in order to generate the artificial bacteria infected mosquito, (the Wolbachia being different from those may being already natural resident in the natural insect population and the Wolbachia inducing cytoplasmic incompatibility (CI) if males of the strain breeding with females from natural population of the insect) wherein the short presence of the additive antibiotic substance in the life cycle of the Tra-2 RNAi strain allowing Wolbachia to be safely maintained in the insect strains; releasing the maleness Tet-on+Tra-2 RNAi/Wolbachia progeny of the strain into the natural population of the species not requiring a sexing step to remove females and achieving population control through breeding with wild females and partially or fully sterilizing them by CI;