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Indel Detection following CRISPR/Cas9 Mutagenesis using High-resolution Melt Analysis in the Mosquito Aedes aegypti
Published on: September 10, 2021
Irradiation induced inversions suppress recombination between the M locus and morphological markers in Aedes aegypti
Antonios A Augustinos1,2, Muhammad Misbah-Ul-Haq3,4, Danilo O Carvalho3
1Insect Pest Control Laboratory, Joint FAO/IAEA Programme of Nuclear Techniques in Food and Agriculture, A-1400, Vienna, Austria. antoniosaugustinos@gmail.com.
Background:
Aedes aegypti is the primary vector of arthropod-borne viruses and one of the most widespread and invasive mosquito species. Due to the lack of efficient specific drugs or vaccination strategies, vector population control methods, such as the sterile insect technique, are receiving renewed interest. However, availability of a reliable genetic sexing strategy is crucial, since there is almost zero tolerance for accidentally released females. Development of genetic sexing strains through classical genetics is hindered by genetic recombination that is not suppressed in males as is the case in many Diptera. Isolation of naturally-occurring or irradiation-induced inversions can enhance the genetic stability of genetic sexing strains developed through genetically linking desirable phenotypes with the male determining region.
Results:
For the induction and isolation of inversions through irradiation, 200 male pupae of the 'BRA' wild type strain were irradiated at 30 Gy and 100 isomale lines were set up by crossing with homozygous 'red-eye' (re) mutant females. Recombination between re and the M locus and the white (w) gene (causing a recessive white eye phenotype when mutated) and the M locus was tested in 45 and 32 lines, respectively. One inversion (Inv35) reduced recombination between both re and the M locus, and wand the M locus, consistent with the presence of a rather extended inversion between the two morphological mutations, that includes the M locus. Another inversion (Inv5) reduced recombination only between w and the M locus. In search of naturally-occurring, recombination-suppressing inversions, homozygous females from the red eye and the white eye strains were crossed with seventeen and fourteen wild type strains collected worldwide, representing either recently colonized or long-established laboratory populations. Despite evidence of varying frequencies of recombination, no combination led to the elimination or substantial reduction of recombination.
Conclusion:
Inducing inversions through irradiation is a feasible strategy to isolate recombination suppressors either on the M or the m chromosome for Aedes aegypti. Such inversions can be incorporated in genetic sexing strains developed through classical genetics to enhance their genetic stability and support SIT or other approaches that aim to population suppression through male-delivered sterility.
Insights
Irradiating Aedes aegypti mosquitoes induced inversions that suppress genetic recombination, enhancing the stability of genetic sexing strains. This supports the sterile insect technique for mosquito population control.
Area of Science:
- Entomology
- Genetics
- Vector Biology
Background:
- Aedes aegypti mosquitoes transmit arboviruses and are a major global invasive species.
- Effective control strategies like the sterile insect technique (SIT) require reliable genetic sexing to prevent female releases.
- Genetic recombination in male Aedes aegypti hinders the development of stable genetic sexing strains.
Purpose of the Study:
- To induce and isolate recombination-suppressing inversions in Aedes aegypti.
- To enhance the genetic stability of genetic sexing strains for vector control.
Main Methods:
- Irradiation of male Aedes aegypti pupae to induce inversions.
- Crossing irradiated males with specific mutant strains to test for reduced recombination.
- Screening wild-type strains for naturally occurring recombination-suppressing inversions.
Main Results:
- One induced inversion (Inv35) reduced recombination between the red-eye (re) and male determining (M) locus, and between the white (w) gene and the M locus.
- A second induced inversion (Inv5) reduced recombination between the w gene and the M locus.
- Naturally occurring inversions did not significantly reduce recombination in tested strains.
Conclusions:
- Irradiation is a viable method to induce inversions that suppress recombination in Aedes aegypti.
- These inversions can improve the genetic stability of sexing strains for SIT and other population control methods.

