Honey bee promoter sequences for targeted gene expression

Schulte, Christina; Leboulle, Gérard; Otte, Marianne; Grünewald, Bernd; Gehne, Nora; Beye, Martin

doi:10.1111/imb.12031

Cited by 15 publications

(13 citation statements)

References 61 publications

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“…The offspring of those transformed queens efficiently expressed the transgenic markers from the 6xP3 promoter (31) and the endogenous promoter Am-actin5c (32). We analyzed at least 34 offspring of each [6xP3-rubia] queen and found that all of the offspring that possessed the transgene were also expressing the transgenic marker (43,6,30, and 9% progeny of the different queens).…”

Section: Resultsmentioning

confidence: 99%

Highly efficient integration and expression of piggyBac -derived cassettes in the honeybee ( Apis mellifera )

Schulte

Theilenberg

Müller-Borg

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

116

120

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Significance We report the first to our knowledge genetically engineered honeybees, which are important pollinators and interesting biological models for the study of social and complex behaviors as well as caste and sexual development. This genetic manipulation tool will enable systematic studies of biological processes in an organism building complex societies. We demonstrate highly efficient integration and expression of piggyBac -derived cassettes in the honeybee that make this system applicable to colony-based screening approaches and useful for an average beekeeping facility. This cassette was stably and efficiently transmitted and expressed in progeny by two different promoters, offering the prospect for activation or inhibition of gene functions under conditions of stage- and tissue-specific promoters.

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Section: Resultsmentioning

confidence: 99%

Highly efficient integration and expression of piggyBac -derived cassettes in the honeybee ( Apis mellifera )

Schulte

Theilenberg

Müller-Borg

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

116

120

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“…It could be that systemic RNAi is not readily achievable in honey bees -indeed, one of the only examples of systemic RNAi in honey bees is from an experiment where the dsRNA corresponds to a honey bee virus, rather than the bees' own genes (17). We are therefore pursuing transgenics as a method of manipulating gene expression in honey bees (18), with the ultimate goal of determining a relationship between OBP expression and death or disease odor detection.…”

Section: Discussionmentioning

confidence: 99%

Results of follow-up experiments to “Odorant cues linked to social immunity induce lateralized antennal stimulation in honey bees (Apis mellifera L.)”

McAfee

Collins

Foster

2017

Preprint

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In 2017, we published the paper "Odorant cues linked to social immunity induce lateralized antennal stimulation in honey bees (Apis mellifera L.)" in Scientific Reports. Since then, we have performed three follow-up experiments which have either negative or contradictory results. Previously, we used electrophysiology to show that hygienic bees displayed significantly higher sensitivity to β-ocimene when stimulated via their left antennae compared to their right. We repeated this assay using worker honey bees from a single hygienic colony and found, to our surprise, that the right antennae elicited higher sensitivity. We also previously attempted to identify a molecular basis for lateralization by using mass spectrometry-based proteomics to compare left and right antennal proteomes. Of the 1,845 proteins, none were differentially expressed. Here, we repeated this experiment but employed orthogonal peptide fractionation to increase proteome coverage to 3,114 proteins; however, still none were differentially expressed. Finally, we attempted to manipulate gene expression of a key antennal odorant binding protein linked to hygienic behaviour (OBP18) using RNA interference via antenna microinjection. We were not able to achieve long-lasting OBP18 knock-down, but comparing the proteomes of untreated, mock dsRNA-treated and OBP18 dsRNA-treated worker antennae revealed numerous off-target effects of the act of injecting alone. By openly reporting this data, we hope to set an example for information transparency.

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“…Robinson et al (2000) attempted to transfect linearized plasmid mixed with sperm into fertilized eggs by the artificial insemination of virgin queens, and reported that the external DNA was propagated for at least three generations, although integration of the transfected DNA into the genome was not detected [47]. Kunieda et al (2004) and Schulte et al (2013) used electroporation to transfect a plasmid into the honey bee brain [48,49]. They confirmed the expression of the external gene ( green fluorescent protein : GFP ) in the brains of transfected bees by immunoblotting or immunohistochemistry using an anti-GFP antibody [48,49].…”

Section: Genetic Methods Applied To the Honey Beementioning

confidence: 99%

Genetics in the Honey Bee: Achievements and Prospects toward the Functional Analysis of Molecular and Neural Mechanisms Underlying Social Behaviors

Kohno

Kubo

2019

Insects

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The European honey bee is a model organism for studying social behaviors. Comprehensive analyses focusing on the differential expression profiles of genes between the brains of nurse bees and foragers, or in the mushroom bodies—the brain structure related to learning and memory, and multimodal sensory integration—has identified candidate genes related to honey bee behaviors. Despite accumulating knowledge on the expression profiles of genes related to honey bee behaviors, it remains unclear whether these genes actually regulate social behaviors in the honey bee, in part because of the scarcity of genetic manipulation methods available for application to the honey bee. In this review, we describe the genetic methods applied to studies of the honey bee, ranging from classical forward genetics to recently developed gene modification methods using transposon and CRISPR/Cas9. We then discuss future functional analyses using these genetic methods targeting genes identified by the preceding research. Because no particular genes or neurons unique to social insects have been found yet, further exploration of candidate genes/neurons correlated with sociality through comprehensive analyses of mushroom bodies in the aculeate species can provide intriguing targets for functional analyses, as well as insight into the molecular and neural bases underlying social behaviors.

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Honey bee promoter sequences for targeted gene expression

Cited by 15 publications

References 61 publications

Highly efficient integration and expression of piggyBac -derived cassettes in the honeybee ( Apis mellifera )

Highly efficient integration and expression of piggyBac -derived cassettes in the honeybee ( Apis mellifera )

Results of follow-up experiments to “Odorant cues linked to social immunity induce lateralized antennal stimulation in honey bees (Apis mellifera L.)”

Genetics in the Honey Bee: Achievements and Prospects toward the Functional Analysis of Molecular and Neural Mechanisms Underlying Social Behaviors

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