The pantry moth, a promising laboratory system for functional genomics in Lepidoptera
Vertical Divider
Plodia interpunctella rearing procedure
(made with Biorender)
The enormous diversity of lepidopteran insects (butterflies and moths) makes their an exciting lineage for exploring new biological questions, but the lack of tractable laboratory system for routine genetic analysis has hindered research. We are developing the pantry moth (Plodia interpunctella) as a laboratory system for discovery in lepidopteran genomes, and hope to open new avenues of research about the physiology, immunology, reproduction, developmental genetics, and evolution of an insect order of enormous diversity that includes numerous species of economic and ecological importance. In addition of being an important pest of stored food across the world, this organism is tailored for laboratory genetics due to a unique suit of reproductive and rearing features:
- ease and low-cost of maintenance
- injectable eggs
- resistance to inbreeding
- successful CRISPR targeting
- an available genome sequence
- translucent stages and tissues that facilitate the screening of fluorescent markers.
Funding : NSF IOS-1923147
EDGE CT: Precise genome editing in a lepidopteran insect tailored for stable transformation (2019-2024)
Vertical Divider
The Plodia Manual (GoogleDoc)
|
CRISPR report : knockouts and
|
Transposon-mediated transgenesis report: hyPBase 3xP3 lines
|
Rearing and microinjection overview
Vertical Divider
Plodia lab rearing and injection setup
(A) Mated Plodia adults in a mass-rearing container.
(B) Injection of CO2 through the container vent.
(C) Transfer into an oviposition jar.
(D-G) Egg transfer into a metallic cup.
(H-J) Inoculation of synchronized embryos in a vented, escape proof container.
(J) Stock at the fifth instar larval stage. Wandering larvae outside of the diet are looking for a pupation site.
(K) Egg injection with microcapillary needles carrying CRISPR or transgenesis reagents.
(L) Microinjection setup.
(A) Mated Plodia adults in a mass-rearing container.
(B) Injection of CO2 through the container vent.
(C) Transfer into an oviposition jar.
(D-G) Egg transfer into a metallic cup.
(H-J) Inoculation of synchronized embryos in a vented, escape proof container.
(J) Stock at the fifth instar larval stage. Wandering larvae outside of the diet are looking for a pupation site.
(K) Egg injection with microcapillary needles carrying CRISPR or transgenesis reagents.
(L) Microinjection setup.
Embryo microinjection workflow
(A) Egg collection from an oviposition jar.
(B) Injection pad using cell culture dish lid.
(C) Egg orientation on parafilm. Water droplets are also added next to the eggs to allow the occasional flushing of the microinjection needle.
(D) Microinjection with glass needle.
(E) Sealing with glue.
(A) Egg collection from an oviposition jar.
(B) Injection pad using cell culture dish lid.
(C) Egg orientation on parafilm. Water droplets are also added next to the eggs to allow the occasional flushing of the microinjection needle.
(D) Microinjection with glass needle.
(E) Sealing with glue.
