A practical guide to CRISPR/Cas9 genome editing in Lepidoptera

Zhang, Linlin; Reed, Robert D.

doi:10.1101/130344

Cited by 17 publications

(19 citation statements)

References 27 publications

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“…S2). The specificity of sgRNA sequence was examined against H. armigera genome using the CRISPR/Cas system (Cas9/gRNA) off‐targeter (CasOT, http://casot.cbi.pku.edu.cn), and the off‐target sites with relatively high risk of mutations were experimentally evaluated …”

Section: Methodsmentioning

confidence: 99%

NPC1b as a novel target in controlling the cotton bollworm, Helicoverpa armigera

Zheng

Yue

Kuang

et al. 2020

Pest Management Science

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BACKGROUND: Insects cannot synthesize sterols and must acquire them from food. The mechanisms underlying how insects uptake dietary sterols are largely unknown except that NPC1b, an integral membrane protein, has been shown to be responsible for dietary cholesterol uptake in Drosophila melanogaster. However, whether NPC1b orthologs in other insect species, particularly the economically important pests, function similarly remains to be determined. RESULTS:In this study, we characterized the function of NPC1b in Helicoverpa armigera, a global pest that causes severe yield losses to many important crops. Limiting dietary cholesterol uptake to insects significantly inhibited food ingestion and weight gain. Compared to the wild-type H. armigera, the CRISPR/Cas9-edited NPC1b mutant larvae were incapable of getting adequate cholesterol and died in their early life stage. Gene expression profile and in situ hybridization analyses indicated that NPC1b was mainly expressed in the midgut where dietary cholesterol was absorbed. Expression of NPC1b was also correlated with the feeding life stages and was especially upregulated during early larval instars. Protein-ligand docking and sequence similarity analyses further demonstrated that NPC1b proteins of lepidopteran insects shared a relatively conserved cholesterol binding region, NPC1b_NTD, which, however, was highly divergent from bees-derived sequences.CONCLUSION: NPC1b was crucial for dietary cholesterol uptake and growth of H. armigera, and therefore could serve as an insecticide target for the development of a novel pest-management approach to control this economically significant insect pest with little off-target effect on bees and sterol-autotrophic animals.

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Section: Methodsmentioning

confidence: 99%

NPC1b as a novel target in controlling the cotton bollworm, Helicoverpa armigera

Zheng

Yue

Kuang

et al. 2020

Pest Management Science

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“…ABC transporters play a key role in the biosynthesis of insect chromes, as has been demonstrated in the genetic models D. melanogaster , B. mori , Helicoverpa armigera and Tribolium castaneum (Ewart et al, ; Tatematsu et al, ; Grubbs et al, ; Khan et al, ). These model species, especially the lepidopteran models, are highly useful in terms of their suitability for CRISPR/Cas9‐based genome editing (Zhang & Reed, ). This method is of great interest now that there are candidate genes to test, which are already known to work in the ommochrome pathway or in ommochromasome biogenesis in other species.…”

Section: Ommochromes Within the Cell: The Ommochromasome Life Cyclementioning

confidence: 99%

“…More recently, developmental biologists have used ommochromes of butterfly wings to tackle the mechanisms and the diversity of colour patterning (Sekimura & Nijhout, ). Today, ommochromes are also involved in transcriptomic studies of grasshoppers, butterflies, damselflies and spiders (Croucher et al, ; Chauhan et al, ; Connahs, Rhen & Simmons, ; Qiu et al, ; Wang et al, ), as well as in clustered regular interspaced short palindromic repeats/CRISPR‐associated protein 9 (CRISPR/Cas9)‐mediated genome editing (Khan, Reichelt & Heckel, ; Xue et al, ; Zhang & Reed, ). Unfortunately, biochemical knowledge did not keep pace with this increase in functional studies.…”

Section: Introductionmentioning

confidence: 99%

Ommochromes in invertebrates: biochemistry and cell biology

2018

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Ommochromes are widely occurring coloured molecules of invertebrates, arising from tryptophan catabolism through the so-called Tryptophan → Ommochrome pathway. They are mainly known to mediate compound eye vision, as well as reversible and irreversible colour patterning. Ommochromes might also be involved in cell homeostasis by detoxifying free tryptophan and buffering oxidative stress. These biological functions are directly linked to their unique chromophore, the phenoxazine/phenothiazine system. The most recent reviews on ommochrome biochemistry were published more than 30 years ago, since when new results on the enzymes of the ommochrome pathway, on ommochrome photochemistry as well as on their antiradical capacities have been obtained. Ommochromasomes are the organelles where ommochromes are synthesised and stored. Hence, they play an important role in mediating ommochrome functions. Ommochromasomes are part of the lysosome-related organelles (LROs) family, which includes other pigmented organelles such as vertebrate melanosomes. Ommochromasomes are unique because they are the only LRO for which a recycling process during reversible colour change has been described. Herein, we provide an update on ommochrome biochemistry, photoreactivity and antiradical capacities to explain their diversity and behaviour both in vivo and in vitro. We also highlight new biochemical techniques, such as quantum chemistry, metabolomics and crystallography, which could lead to major advances in their chemical and functional characterisation. We then focus on ommochromasome structure and formation by drawing parallels with the well-characterised melanosomes of vertebrates. The biochemical, genetic, cellular and microscopic tools that have been applied to melanosomes should provide important information on the ommochromasome life cycle. We propose LRO-based models for ommochromasome biogenesis and recycling that could be tested in the future. Using the context of insect compound eyes, we finally emphasise the importance of an integrated approach in understanding the biological functions of ommochromes.

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“…While now there is a consensus to view species as natural entities delineated by multiple evidences resulting from differentiation of lineages through speciation [11], species boundaries are often much harder to discern especially in a Lepidoptera collection because specimens are sampled across vast geographical areas and through time [4,11,13,18,21]. Furthermore, the order Lepidoptera challenges species boundaries for regular potential or often occurring interbreeding [34]. Despite this, a species is a central concept in biology, conservation, legislation, and trade and therefore it has a particular social relevance to the museum collection and to the society.…”

Section: Delineating Identifying and Describing The Taxamentioning

confidence: 99%

Lepidoptera Collection Curation and Data Management

Prins¹

2017

Lepidoptera

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The collections of Lepidoptera often serve as foundational basis for a wide range of biological, ecological, and climate science disciplines. Species identification and higher taxa delimitation based on collection specimens and especially, on types test scientific hypotheses, provide multiple types of evidence for a broad range of users. Curation and data management approaches applied in Lepidoptera collections benefit greatly from many newly developed information techniques, which link and integrate data. Mostly attention is focused on clean verified collection and taxonomic literature mining data to obtain correct species-group and higher taxa names, as well as reliable data on the distribution of Lepidoptera and their trophic interactions. Collection creation and management became a subject of natural sciences itself. The chapter provides a historic overview on collection creation and curation together with a short discussion on collection goals and purposes. The creation of a virtual collection based on interlinked data is emphasized. Information science and data management tools became very important in Lepidoptera collection curation. The complexity of techniques and computing tools used in taxonomy and the increase in the amount of data that can be obtained by collectionbased disciplines make it necessary to automate data gathering, manipulation, analysis, and visualization processes.

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A practical guide to CRISPR/Cas9 genome editing in Lepidoptera

Cited by 17 publications

References 27 publications

NPC1b as a novel target in controlling the cotton bollworm, Helicoverpa armigera

NPC1b as a novel target in controlling the cotton bollworm, Helicoverpa armigera

Ommochromes in invertebrates: biochemistry and cell biology

Lepidoptera Collection Curation and Data Management

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