Insect Transgenesis and the Sterile Insect Technique

Schetelig, Marc F.; Wimmer, Ernst A.

doi:10.1007/978-90-481-9641-8_9

Cited by 13 publications

(14 citation statements)

References 168 publications

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“…aegypti genome, showed positive results and proved the efficient vertical transmission of the reporter gene. As expected, based on previous reports (Schetelig and Wimmer, 2011), these results indicate that the ECFP protein showed no detrimental effect on the adult tissues of Ae. aegypti, as we speculate it happened with the eGFP and the scorpine on adult tissues of An.…”

Section: Discussionsupporting

confidence: 92%

Genetic transformation of mosquitoes by microparticle bombardment

Lule-Chávez

Carballar-Lejarazú

Cabrera-Ponce

et al. 2020

Insect Molecular Biology

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Mosquitoes constitute the major living beings causing human deaths in the world. They are vectors of malaria, yellow fever, dengue, zika, filariases, chikungunya, among other diseases. New strategies to control/eradicate mosquito populations are based on newly developed genetic manipulation techniques. However, genetic transformation of mosquitoes is a major technical bottleneck due to low efficiency, the need of sophisticated equipment, and highly trained personnel. The present report shows the transgenerational genetic transformation of Aedes aegypti, using the particle inflow gun (PIG), by integrating the ecfp gene in the AAEL000582 mosquito gene with the CRISPR‐Cas9 technique, achieving a mean efficiency of 44.5% of bombarded individuals (G0) that showed ECFP expression in their tissues, and a mean of 28.5% transformation efficiency measured on G1 individuals. The same transformation technique was used to integrate the egfp/scorpine genes cloned in the Minos transposon pMinHygeGFP into the Anopheles albimanus genome, achieving a mean efficiency of 43.25% of bombarded individuals (G0) that showed EGFP expression in their tissues. Once the technique was standardized, transformation of Ae. aegypti neonate larvae and An. albimanus eggs was achieved when exposed to gold microparticle bombardment. Integration of genes and heterologous protein expression were confirmed by PCR, sequencing, fluorescent microscopy, mass spectrometry, Western blot and dot blot analyses. Transgenerational inheritance of the transgenes was observed only on Ae. aegypti, as all transformed An. albimanus individuals died at the pupal stage of the G0 generation.

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Section: Discussionsupporting

confidence: 92%

Genetic transformation of mosquitoes by microparticle bombardment

Lule-Chávez

Carballar-Lejarazú

Cabrera-Ponce

et al. 2020

Insect Molecular Biology

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“…Biotechnological engineering of insects makes novel approaches possible to efficiently mark insects as well as selectively produce vigorous and potent sterile males, which are generated by conditional male reproductive sterility in combination with conditional female lethality. This will improve efficacy and widen applicability to further insect pest species [ 7 , 8 ]. To minimize the concerns coupled with the release of transgenic organisms, SIT programs are actually ideal, as the sterility of the released males will serve as a biological safety mechanism for containment as it impedes the spread of transgenes and allows for a safe deployment [ 9 , 10 ].…”

Section: Introductionmentioning

confidence: 99%

Perspective on the combined use of an independent transgenic sexing and a multifactorial reproductive sterility system to avoid resistance development against transgenic Sterile Insect Technique approaches

Eckermann¹,

Dippel²,

KaramiNejadRanjbar³

et al. 2014

BMC Genet

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BackgroundThe Sterile Insect Technique (SIT) is an accepted species-specific genetic control approach that acts as an insect birth control measure, which can be improved by biotechnological engineering to facilitate its use and widen its applicability. First transgenic insects carrying a single killing system have already been released in small scale trials. However, to evade resistance development to such transgenic approaches, completely independent ways of transgenic killing should be established and combined.PerspectiveMost established transgenic sexing and reproductive sterility systems are based on the binary tTA expression system that can be suppressed by adding tetracycline to the food. However, to create 'redundant killing' an additional independent conditional expression system is required. Here we present a perspective on the use of a second food-controllable binary expression system - the inducible Q system - that could be used in combination with site-specific recombinases to generate independent transgenic killing systems. We propose the combination of an already established transgenic embryonic sexing system to meet the SIT requirement of male-only releases based on the repressible tTA system together with a redundant male-specific reproductive sterility system, which is activated by Q-system controlled site-specific recombination and is based on a spermatogenesis-specifically expressed endonuclease acting on several species-specific target sites leading to chromosome shredding.ConclusionA combination of a completely independent transgenic sexing and a redundant reproductive male sterility system, which do not share any active components and mediate the induced lethality by completely independent processes, would meet the 'redundant killing' criteria for suppression of resistance development and could therefore be employed in large scale long-term suppression programs using biotechnologically enhanced SIT.

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“…Synthetic regulatory elements containing multiple adjacent binding sites for a small number of transcriptional activators have not been used so far in published gene drive constructs but they might in the future. In general, such synthetic pieces tend to be more universal than endogenous regulatory elements (Schetelig and Wimmer 2011) . For example, the Drosophila-derived 3xP3 artificial element drives expression in the eyes not only in insects but also in arthropods (Pavlopoulos and Averof 2005) and planarians (Gonzalez-Estevez et al 2003) .…”

Section: Probability That the Guide Rna And Cas9 Genes Are Expressed mentioning

confidence: 99%

Evaluating the Probability of CRISPR-based Gene Drive Contaminating Another Species

Courtier‐Orgogozo

Danchin

Gouyon

et al. 2019

Preprint

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The probability D that a given CRISPR-based gene drive element contaminates another, non-target species can be estimated by the following Drive Risk Assessment Quantitative Estimate (DRAQUE) Equation: D = ( hyb+transf).express.cut.flank.immune.nonextinct with hyb = probability of hybridization between the target species and a non-target species transf = probability of horizontal transfer of a piece of DNA containing the gene drive cassette from the target species to a non-target species (with no hybridization) express = probability that the Cas9 and guide RNA genes are expressed cut = probability that the CRISPR-guide RNA recognizes and cuts at a DNA site in the new host flank = probability that the gene drive cassette inserts at the cut site immune = probability that the immune system does not reject Cas9 -expressing cells nonextinct = probability of invasion of the drive within the populationWe discuss and estimate each of the seven parameters of the equation, with particular emphasis on possible transfers within insects, and between rodents and humans. We conclude from current data that the probability of a gene drive cassette to contaminate another species is not insignificant. We propose strategies to reduce this risk and call for more work on estimating all the parameters of the formula.

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Insect Transgenesis and the Sterile Insect Technique

Cited by 13 publications

References 168 publications

Genetic transformation of mosquitoes by microparticle bombardment

Genetic transformation of mosquitoes by microparticle bombardment

Perspective on the combined use of an independent transgenic sexing and a multifactorial reproductive sterility system to avoid resistance development against transgenic Sterile Insect Technique approaches

Evaluating the Probability of CRISPR-based Gene Drive Contaminating Another Species

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