Team:Mingdao/Applied Design

To demonstrate our pathogen blood testing system on a real adult mosquito, we collaborated with NCHU_Taichung iGEM Team 2018 from Taiwan and conducted an experiment of DNA microinjection into adult female mosquito, Aedes aegypti.

In addition to this experiment, we also visited the Mosquito Lab of Prof. Wu-Chun Tu in National Chung Hsing University, Taiwan. We presented our idea and got the valuable feedback from him. Moreover, we learned the physiology and behavior of mosquitoes when sucking and attracted by human blood from Dr. Hau-you Tzeng. The knowledge we learned was turned into our prototype design.

EXPERIMENT at NCHU

↓ Make microinjection needles by pulling glass capillary tubes
↓ Anesthetize the adult mosquitoes on ice
↓ Fill the needle with the DNAs in Cellfectin II reagent with E. coli or not
↓ Inject the DNA by inserting into the thorax of the mosquito.
↓ Recover the mosquito for 2 days

RESULT

The mosquito injected with DNA plus E. coli showed fluorescence signal in Gel Imaging System and Blue LED Box after 2 days of DNA microinjection. This result further proved our concept into a reality.

To make GE mosquito GFP reporter system come to the world, some may be considered: product packaging, result reading device and protocol, safety, as well as product marketing.

We developed two products as one set for our reporter and reading system:

1. KISSING BOX^TM: GE mosquitoes kept in the box for sucking blood

2. BLUE LED BOX: a scanner box equipped with blue led for observing the green fluorescence signal.

We made a matchbox-size KISSING BOX^TM with a clear acrylic sheet (as a transparent window) on the top and a cover of fine metal mesh in the bottom. The box can carry as many as 20 mosquitoes which can suck blood through the mesh.

The SCANNER can load a KISSING BOX^TM inside.

The SCANNER equipped with blue LED which can excite the green fluorescence of GFP as shown here with fluorescein in a PCR tube.

You can see glowing mosquitoes sucking fluorescein through the transparent window in KISSING BOX^TM.

The SCANNER BOX with KISSING BOX^TM is light and portable which can be held with one hand. The operation is simple and easy-to-use without any profession instructions. Our product is suitable for point-of-care use including poor-educated and limited-resource countries.

Compared to other current methods, the KISSING BOX^TM provides a novel way solving difficulties of accuracy, windows period of virus infection, measuring time and place. The KISSING BOX^TM is developed for point-of-care use and affordable to the public.

Safety is more than everything. The product will bring to you with wingless GE mosquitoes either with wings cut manually or genetically modified by knocking out a wing development gene, vestigial.