Fig. 4: Demonstration of CRISPR in G. mellonella.
From: PiggyBac-mediated transgenesis and CRISPR–Cas9 knockout in the greater wax moth, Galleria mellonella
a, Bmhsp90:GFP/3xP3:DsRed G0 larva I–V have an expression cassette inserted into their genome resulting in the expression of eGFP visible in somatic unmelanized tissue and the expression of dsRed in both eye (solid arrow) and neural (dashed arrow) tissue. Larva I is the G1 offspring of individuals of the same transgenic line as larvae II–V, but injected with a Cas9/sgRNA ribonucleoprotein targeting the eGFP gene, causing a heritable loss of function in the eGFP gene, but retaining the eye-specific dsRed expression. Larvae II–V are of the parent strain and were not injected with Cas9/sgRNA. b, Mutations within the coding sequences of 12 different GFP-negative G1 individuals. The G1 larvae are offspring from individuals crosses of Bmhsp90:GFP/3xP3:DsRed G0s injected with an sgRNA targeting the N terminus of the eGFP sequence and crossed to WT mates. The top line is the native eGFP sequence, with the sequence complementary to the sgRNA highlighted in green and the PAM sequence highlighted in red.
