Genetically Encoded CRISPR Components Yield Efficient Gene Editing in the Invasive Pest <i>Drosophila suzukii</i>

Kandul, Nikolay P.; Belikoff, Esther J.; Liu, Junru; Buchman, Anna; Li, Fang; Yamamoto, Akihiko; Yang, Ting; Shriner, Isaiah; Scott, Maxwell J.; Akbari, Omar S.

doi:10.1089/crispr.2021.0032

Cited by 16 publications

(19 citation statements)

References 61 publications

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“…Without the tools that we exploited in engineering SSIMS, such as multiple genetic markers and balancer chromosomes, engineering the SSIMS genotype in non-model insects will require more tedious molecular characterization of offspring produced during strain construction. However, we are encouraged by recent publications highlighting new genetic tools for non-model organisms ( Kandul et al, 2021 ). Also, the molecular components needed to engineer SSIMS have been shown to work in diverse organisms.…”

Section: Discussionmentioning

confidence: 99%

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Upadhyay

Feltman

Sychla

et al. 2022

eLife

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Engineered Genetic Incompatibility (EGI) is a method to create species-like barriers to sexual reproduction. It has applications in pest control that mimic Sterile Insect Technique when only EGI males are released. This can be facilitated by introducing conditional female-lethality to EGI strains to generate a sex-sorting incompatible male system (SSIMS). Here, we demonstrate a proof of concept by combining tetracycline-controlled female lethality constructs with a pyramus-targeting EGI line in the model insect Drosophila melanogaster. We show that both functions (incompatibility and sex-sorting) are robustly maintained in the SSIMS line and that this approach is effective for population suppression in cage experiments. Further we show that SSIMS males remain competitive with wild-type males for reproduction with wild-type females, including at the level of sperm competition.

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Section: Discussionmentioning

confidence: 99%

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Upadhyay

Feltman

Sychla

et al. 2022

eLife

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“…Cas9-mediated cleavage of genomic DNA followed by homology-dependent repair using the injected plasmid DNA as template should lead to integration of the FL19 construct at the targeted location ( Wang et al, 2015 ; Davis et al, 2018 ). These experiments would be facilitated by using the lines that express Cas9 in the germline that we described recently ( Kandul et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Additional Drosophila suzukii Male-Only Strains Generated Through Remobilization of an FL19 Transgene

Yamamoto

Yadav

Scott

2022

Front. Bioeng. Biotechnol.

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Drosophila suzukii (D. suzukii) (Matsumura, 1931; Diptera: Drosophilidae), also known as spotted wing Drosophila, is a worldwide pest of fruits with soft skins such as blueberries and cherries. Originally from Asia, D. suzukii is now present in the Americas and Europe and has become a significant economic pest. Growers largely rely on insecticides for the control of D. suzukii. Genetic strategies offer a species-specific environmentally friendly way for suppression of D. suzukii populations. We previously developed a transgenic strain of D. suzukii that produced only males on a diet that did not contain tetracycline. The strain carried a single copy of the FL19 construct on chromosome 3. Repeated releases of an excess of FL19 males led to suppression of D. suzukii populations in laboratory cage trials. Females died as a consequence of overexpression of the tetracycline transactivator (tTA) and tTA-activated expression of the head involution defective proapoptotic gene. The aim of this study was to generate additional male-only strains that carried two copies of the FL19 transgene through crossing the original line with a piggyBac jumpstarter strain. Males that carried either two chromosome 3 or a singleX-linked transgene were identified through stronger expression of the red fluorescent protein marker gene. The brighter fluorescence of the X-linked lines was likely due to dosage compensation of the red fluorescent protein gene. In total, four X-linked lines and eleven lines with two copies on chromosome 3 were obtained, of which five were further examined. All but one of the strains produced only males on a diet without tetracycline. When crossed with wild type virgin females, all of the five two copy autosomal strains examined produced only males. However, the single copy X-linked lines did not show dominant female lethality. Five of the autosomal lines were further evaluated for productivity (egg to adult) and male competition. Based on these results, the most promising lines have been selected for future population suppression experiments with strains from different geographical locations.

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“…( Kandul et al, 2022;Li et al, 2021;Schetelig et al, 2021). In this regard, progress has been made with strains developed that express Cas9 in the germline (Kandul et al, 2021) and under heat shock conditions (Yan et al, 2021). To facilitate transgenesis, lines that express piggyBac transposase (Chu et al, 2018) and phiC31 recombinase (Ahmed et al, 2020) have been established and characterised.…”

Section: Fecundity and Fertility Of White Cinnabar And Sepia Strainsmentioning

confidence: 99%

“…For example, Morgan (1910) discovered the white mutant over 110 years ago; and even today, it is one of the most useful tools in the fruit fly genetics research. For D. suzukii , we previously reported the isolation of mutant alleles of the X‐linked white and yellow genes, which produce flies with white eyes and yellow bodies, respectively (Kandul et al, 2021; Li & Scott, 2016). The white strains have not been as useful as for D. melanogaster because of the sterility of white‐eyed D. suzukii males (Li & Scott, 2016; Yan et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

“…The clustered regularly interspaced short palindromic repeats (CRISPRs)/CRISPR‐associated protein 9 (Cas9) system was used to obtain D. suzukii white and yellow mutants (Kalajdzic & Schetelig, 2017; Kandul et al, 2021; Li & Scott, 2016). CRISPR/Cas9 was first identified as part of a bacterial immune defence system (Barrangou & Horvath, 2017; Wang & Doudna, 2023).…”

Section: Introductionmentioning

confidence: 99%

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Expansion of the genetic toolbox for manipulation of the global crop pest Drosophila suzukii: Isolation and assessment of eye colour mutant strains

Yadav,

Asokan,

Yamamoto

et al. 2023

Insect Molecular Biology

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Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), commonly called spotted wing Drosophila, is an important agricultural pest recognised worldwide. D. suzukii is a pest of soft‐skinned fruits as females can lay eggs in ripening fruit before harvest. While strains for genetic biocontrol of D. suzukii have been made, the development of transgenic D. suzukii strains and their further screening remain a challenge partly due to the lack of phenotypically trackable genetic‐markers, such as those widely used with the model genetic organism D. melanogaster. Here, we have used CRISPR/Cas9 to introduce heritable mutations in the eye colour genes white, cinnabar and sepia, which are located on the X, second and third chromosomes, respectively. Strains were obtained, which were homozygous for a single mutation. Genotyping of the established strains showed insertion and/or deletions (indels) at the targeted sites. A strain homozygous for mutations in cinnabar and sepia showed a pale‐yellow eye colour at eclosion but darkened to a sepia colour after a week. The fecundity and fertility of some of the cinnabar and sepia strains were comparable with the wild type. Although white mutant males were previously reported to be sterile, we found that sterility is not fully penetrant and we have been able to maintain white‐eyed strains for over a year. The cinnabar, sepia and white mutant strains developed in this study should facilitate future genetic studies in D. suzukii and the development of strains for genetic control of this pest.

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Genetically Encoded CRISPR Components Yield Efficient Gene Editing in the Invasive Pest Drosophila suzukii

Cited by 16 publications

References 61 publications

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Evaluation of Additional Drosophila suzukii Male-Only Strains Generated Through Remobilization of an FL19 Transgene

Expansion of the genetic toolbox for manipulation of the global crop pest Drosophila suzukii: Isolation and assessment of eye colour mutant strains

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