Male-sterile Anopheles gambiae mosquitoes | BCH-LMO-SCBD-260489 | Living Modified Organism | Biosafety Clearing-House

Loading...
Living Modified Organism (LMO)
  |  
Decisions on the LMO Risk Assessments  
last updated: 30 May 2022
Living Modified Organism identity
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.
Male-sterile Anopheles gambiae mosquitoes
EN
Ag(DSM)2
No
The African malaria mosquito (Anopheles gambiae) was modified for male sterility and to have a fluorescent marker. For sterility, the modified mosquito expresses a homing endonuclease gene (Physarum polycephalum intron-encoded endonuclease; I-PpoI), which cuts a ribosomal DNA sequence on the X chromosome during male spermatogenesis. However, all spermatozoa, irrespectively of the inheritance of the gene cassettes, carried a substantial amount of I-PpoI protein. Thus, upon mating and fertilization of the egg, the maternal X chromosome is also cut due to the activity of I-PpoI delivered from the sperm, leading to developmental arrest of embryos and thereby resulting in no viable progeny. Thus, the destruction of the X chromosomes renders the male mosquitoes sexually sterile and unable to father viable progeny as a result of the I-PpoI activity on the X chromosome. In addition, the I-PpoI endonuclease is fused to Aequorea victoria enhanced green fluorescent protein, which was used to verify expression of I-PoI in the testes. The modified mosquito also expresses Discosoma sp. red fluorescent protein (DsRed2), which acts as a marker, being expressed in the eyes of the modified mosquitoes.
EN
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
  • BCH-ORGA-SCBD-260392-1 Organism Anopheles gambiae (African malaria mosquito, Mosquito, Malaria mosquito, ANOGA)
    Insects
Anopheles gambiae G3 strain eggs from BEIresources database
EN
Characteristics of the modification process
pBac{3xP3-DsRed}b2-eGFPI-PpoI
EN
  • Microinjection
 
0.000 kb
 
 
0.000 kb
 
 
0.000 kb
 
 
0.000 kb
 
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-115246-2 piggyBac | Trichoplusia ni - Cabbage looper, Cabbage looper moth, Cabbage plusia, Common cabbage looper, Lettuce looper
    Transposon
  • BCH-GENE-SCBD-260479-1 3xP3 promoter | Drosophila melanogaster (Common Fruit Fly)
    Promoter
  • BCH-GENE-SCBD-101476-6 DsRed2 Fluorescent Protein | Discosoma sp. (Coral anemones, Sea anemones)
    Protein coding sequence | Changes in quality and/or metabolite content (Pigmentation / Coloration)
  • BCH-GENE-SCBD-114748-3 SV40 poly-adenylation signal | Macaca mulatta polyomavirus 1 (SV40, Simian vacuolating virus 40, simian virus 40, Rhesus macaque polyomavirus)
    Terminator
  • BCH-GENE-SCBD-260481-1 Beta-2 tubulin promoter | Anopheles gambiae (African malaria mosquito, Mosquito, Malaria mosquito, ANOGA)
    Promoter
  • BCH-GENE-SCBD-260488-1 Enhanced green fluorescent protein | Aequorea victoria (Crystal Jellyfish, Water Jellyfish, AEQVI)
    Protein coding sequence | Changes in quality and/or metabolite content (Pigmentation / Coloration),Selectable marker genes and reporter genes
  • BCH-GENE-SCBD-260487-4 Intron-encoded endonuclease | Physarum polycephalum (Plasmodial slime molds, Slime mold, PHYPO)
    Protein coding sequence | Changes in physiology and/or production (Reproduction, Male sterility),Engineered gene drive application (Population suppression)
  • BCH-GENE-SCBD-260483-1 Beta-2 tubulin terminator | Anopheles gambiae (African malaria mosquito, Mosquito, Malaria mosquito, ANOGA)
    Terminator
The modified Anopheles gambiae mosquitoes contain two gene cassettes: Discosoma sp. red fluorescent protein DsRed2 and an Aequorea victoria enhanced green fluorescent protein-Physarum polycephalum intron-encoded endonuclease (eGFP::I-Ppol) fusion protein. The non-autonomous Trichoplusia ni piggyBac transposable element was used to insert the gene cassettes into the germline of the modified mosquitoes.

The DsRed2 coding sequence is under control of the synthetic 3xP3 promoter and the Macaca mulatta polyomavirus 1 SV40 polyadenylation signal. The photoreceptor-specific nature of the promoter restricts expression of DsRed2 to the eyes of the modified mosquito.

The eGFP::I-Ppol coding sequence is under control of an An. gambiae beta-2 tubulin promoter and terminator. Due to the nature of the promoter, expression of the eGFP::I-Ppol is expected to be restricted to male spermatogenesis in the testes. The eGFP protein makes up the N-terminal region of the fusion protein, while I-PpoI is on the C-terminal end.
EN
LMO characteristics
EN
  • Research
Detection method(s)
Modified adult mosquitoes can be detected by the red fluorescence in their eyes due to the presence of the DsRed2 protein.
EN
Additional Information
Anopheles gambiae sensu lato (s.l.) is a species complex, whose members are morphologically indistinguishable in their adult stage but isolated reproductively. They have distinct genetic and behavioural traits. Thus, their identification relies on molecular methods. The main malaria vectors in the complex are An. gambiae, An. coluzzii, and An. arabiensis.
EN
Loading...