Mapping and CRISPR homology-directed repair of a recessive white eye mutation in Plodia moths

Functional genomic tools for emerging model species

“…To exchange one allele for another one must instead rely on the homology-directed repair (HDR) pathway, which occurs less commonly in the cell and thus is less frequent than targeted mutagenesis via the NHEJ pathway. However, the HDR pathway enables allelic replacement experiments, something which is attractive for functional validation of quantitative trait loci (QTLs) [56] and the assessment of ecologically relevant variation, as compared to complete KO of gene function. Recent works have also successfully performed knock-ins by relying on homology-independent repair mechanisms in model organisms (Drosophila [57] and zebrafish [58]), and these may also be of use in emerging model organisms in the future.…”

Efficient hyperactive piggyBac transgenesis in Plodia pantry moths

“…We then co-injected the donor plasmid pBac[3xP3::EGFP; nosO::mScarlet] with a hyPBase mRNA and monitored somatic transformation efficiencies throughout the G 0 generation. In order to facilitate the screening of fluorescence, all experiments were performed in the Pi_wFog white-eyed strain that is devoid of screening pigments in eye tissues and also shows increased larval translucency (Heryanto et al, 2022). Transformed embryos and first instar hatchlings showed ocellar and glial EGFP fluorescence (Figures 2C, D), with 23.7% of injected eggs showing EGFP in 72-h embryos (Table 1).…”

“…Previously, delivery of the original pBase as a helper plasmid into Plodia syncytial embryos resulted in somatic transformation of fluorescent markers, but its efficiency for germline transformation was not reported (Bossin et al, 2007). Here, we extend the assessment of hyPBase transgenesis in Lepidoptera with a focus on the pyralid moth P. interpunctella, a pest of stored foods that is amenable to genome editing and genetic transformation (Bossin et al, 2007;Heryanto et al, 2022). In the current study, we injected an insect codon-optimized hyPBase as an mRNA (Otte et al, 2018) and monitored both the somatic and germline transformation rates of fluorescent markers driven by the 3xP3 promoter, a canonical promoter with strong activity in the ocular and glial tissues in Lepidoptera and other insects (Berghammer et al, 1999;Horn et al, 2002;Thomas et al, 2002).…”

“…Mass egg-laying can be stimulated by exposing their highly fecund females (Mbata, 1985) to CO 2 gas, a property that allows the collection of synchronized embryos within the time frame of the first cell divisions, thus facilitating genetic transformation by microinjection (Dyby and Silhacek, 1997;Bossin et al, 2007). Finally, several genome assemblies and several transcriptomic resources have been published in this species (Harrison et al, 2012;Tang et al, 2017;Roberts et al, 2020;Heryanto et al, 2022;Kawahara et al, 2022).…”

Effect of behavioral conditions on silk characteristics in the Indian meal moth (Plodia interpunctella)

“…Other lepidopterans, such as the luna moth ( Actias luna, Linnaeus, Saturniidae; Chen et al, 2012; Reddy and Yang, 2012), and the Indian meal moth ( Plodia interpunctella, Hübner, Pyralidae; Milutinović et al, 2020; Shim and Lee, 2015) are now being studied as the search for tougher, stronger, or more flexible silks (Yoshioka et al, 2019). While recent studies on P. interpunctella have focused on it as a model organism for genomic study of silk production and evolution (Kawahara et al, 2022), still little is known about physiological properties of silk, variables driving phenotypic plasticity, and how behavioral and environmental variables impact silk production (Harrison et al, 2012; Heryanto et al, 2022; Roberts et al, 2020; Tang et al, 2017).…”