Establishment of highly efficient transgenic system for black soldier fly (Hermetia illucens)

Abstract

The black soldier fly (BSF), Hermetia illucens, is a promising insect for mitigating solid waste problems as its larvae are able to bioconvert organic waste into valuable biomass. We recently reported a high-quality genome assembly of the BSF; analysis of this genome sequence will further the understanding of insect biology and identify genes that can be manipulated to improve efficiency of bioconversion. To enable genetic manipulation of the BSF, we have established the first transgenic methods for this economically important insect. We cloned and identified the ubiquitous actin5C promoter (Hiactin5C-p3k) and 3 endogenous U6 promoters (HiU6:1, HiU6:2, and HiU6:3). The Hiactin5C promoter was used to drive expression of a hyperactive variant of the piggyBac transposase, which exhibited up to 6-fold improvement in transformation rate when compared to the wild-type transposase. Furthermore, we evaluated the 3 HiU6 promoters using this transgenic system. HiU6:1 and HiU6:2 promoters provided the highest knockdown efficiency with RNAi and are thus promising candidates for future Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) development. Overall, our findings provide valuable genetic engineering toolkits for basic research and genetic manipulation of the BSF.

Rapid generation and screening of transgenic black soldier fly (Hermetia illucens)

“…offspring. 12 No data on which individual G0 produced transgenic offspring was recorded in our study due to large group matings and the inability to determine which egg clutches came from which G0 parents. However, if we consider that all fertilised clutches that we collected were only laid from matings between fertile adults, the percentage of those clutches containing transgenic offspring is an indirect measure of the transformation rate.…”

“…The previously reported rate of germline transmission was 10 out of 84 (11.9%), this transformation rate was a measure of the percentage of fertile G0 transgenic adults transmitting the transgene to G1 offspring 12 . No data on which individual G0 produced transgenic offspring was recorded in our study due to large group matings and the inability to determine which egg clutches came from which G0 parents.…”

Enhancing amino acid productivity and profile in black soldier fly larvae through NAT transporter suppression in the excretion system

“…The method we described in this study represents a ground-breaking departure from previous research. Other approaches to enhancing the productivity of protein in BSFL have primarily focused on manipulating growth hormones to increase larval body size and growth rate (Gruber and Melton, 2023; Kou et al ., 2022; Zhan et al ., 2020). In contrast, our study introduces a novel strategy: the regulation of membrane transport within the excretory system to significantly boost the total protein content per individual larva.…”

BSFbase: The comprehensive genomic resource for a natural recycler, the black soldier fly (Hermetia illucens L.)

“…Moreover, the CRISPR/Cas9-based genome editing system has been successfully applied in many non-model systems, providing a powerful way of studying the functions of given genes in focal species (Xu et al, 2019). Efficient gene modification in BSF has also recently been reported (Zhan et al, 2019;Kou et al, 2022), highlighting the possibility of revealing the functions of genes and selecting more economically beneficial strains.…”

“…, 2019; Kou et al. , 2022), highlighting the possibility of revealing the functions of genes and selecting more economically beneficial strains.…”