Somatic transformation efficiencies and expression patterns using the <i>Jc</i>DNV and <i>piggyBac</i> transposon gene vectors in insects

Bossin, Hervé; Furlong, Richard B.; Gillett, J. L.; Bergoin, Max; Shirk, Paul D.

doi:10.1111/j.1365-2583.2006.00693.x

Cited by 17 publications

(52 citation statements)

References 31 publications

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“…By combining PB calling cards with available transposon tools (such as gene traps), one might be able to enrich for transposon insertions in genes of specific pathways for mutagenesis screens. Since piggyBac is active in many different species, including yeast (Mitra et al 2008), insects (Bossin et al 2007), zebrafish (Lobo et al 2006), mouse, and human (Ding et al 2005), the calling card method could enjoy wide use.…”

Section: Discussionmentioning

confidence: 99%

“…By harvesting the transposon calling cards along with their flanking genomic DNA, a genome-wide map of transcription factor binding can be obtained. We harnessed the piggyBac (PB) transposon as a calling card (Cary et al 1989;Ding et al 2005), due to its key advantages over other transposons: it is active in many different species, including yeast (Mitra et al 2008), insects (Bossin et al 2007), zebrafish (Lobo et al 2006), mouse, and human (Ding et al 2005), its transposition efficiency is high, and PB transposase is amenable to addition of proteins to its N terminus (Wu et al 2006;Wilson et al 2007) (Supporting Information, Figure S1). …”

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confidence: 99%

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“Calling Cards” for DNA-Binding Proteins in Mammalian Cells

et al. 2012

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The ability to chronicle transcription-factor binding events throughout the development of an organism would facilitate mapping of transcriptional networks that control cell-fate decisions. We describe a method for permanently recording protein-DNA interactions in mammalian cells. We endow transcription factors with the ability to deposit a transposon into the genome near to where they bind. The transposon becomes a "calling card" that the transcription factor leaves behind to record its visit to the genome. The locations of the calling cards can be determined by massively parallel DNA sequencing. We show that the transcription factor SP1 fused to the piggyBac transposase directs insertion of the piggyBac transposon near SP1 binding sites. The locations of transposon insertions are highly reproducible and agree with sites of SP1-binding determined by ChIP-seq. Genes bound by SP1 are more likely to be expressed in the HCT116 cell line we used, and SP1-bound CpG islands show a strong preference to be unmethylated. This method has the potential to trace transcription-factor binding throughout cellular and organismal development in a way that has heretofore not been possible.

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Section: Discussionmentioning

confidence: 99%

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“Calling Cards” for DNA-Binding Proteins in Mammalian Cells

et al. 2012

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“…I). The limited frequency of expression from foreign promoters has been observed previously in other somatically transformed insects (Bossin et al ., ). None of the microinjected N. vitripennis embryos survived past the first larval instars.…”

Section: Resultsmentioning

confidence: 97%

Functional characterization of the transcriptional regulatory elements of three highly expressed constitutive genes in the jewel wasp, Nasonia vitripennis

Shirk

Furlong

Dolan

et al. 2017

Insect Molecular Biology

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The jewel wasp, Nasonia vitripennis Ashmead (Hymenoptera: Pteromalidae), is an easily reared parasitoid that is providing an ever increasingly malleable model for examining the biology and genetics of Hymenoptera. Utilizing genomic and transcriptome resources, 5' upstream transcriptional regulatory sequences (TREs) from three highly expressed genes were identified and cloned. Criteria for TRE selection included the presence of an adjacent gene 5' of the translation initiation site. One gene was methylated whereas the other two were nonmethylated. Each TRE, heat-shock protein 70 (hsp70), activator of 90 kDa hsp ATPase protein 1 (hsp90A), and lipid storage droplet surface-binding protein 1 (lsdp) was linked with enhanced green fluorescent protein (EGFP) coding sequence and cloned into both pDP9e somatic and piggyBac germline transformation vectors. EGFP expression patterns under control of each TRE were compared with patterns of DsRed fluorescence produced from the transformation vector cassette. Functional activity of each TRE was observed in cultured Spodoptera frugiperda 9 (Sf9) cells and Drosophila melanogaster as well as in N. vitripennis embryos demonstrating that all three sequences had functional transcriptional regulatory activity in three different insect orders. Identification and functional characterization of these three TREs will provide critical and necessary resources for further genetic analyses of N. vitripennis, Hymenoptera and other insects.

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“…Therefore, inclusion of cultured honeybee cells [64] may provide a more attractive and complete feeding medium. Because the cultured insect cells can be genetically transformed [65, 66], supplementation of the feeding medium can provide an additional means of introducing various constructs that will produce materials that can be tested for control of the mites.…”

Section: Discussionmentioning

confidence: 99%

A feeding protocol for delivery of agents to assess development in Varroa mites

2017

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A novel feeding protocol for delivery of bio-active agents to Varroa mites was developed by providing mites with honey bee larva hemolymph supplemented with cultured insect cells and selected materials delivered on a fibrous cotton substrate. Mites were starved, fed on treated hemolymph to deliver selected agents and then returned to bee larvae. Transcript levels of two reference genes, actin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), as well as for nine selected genes involved in reproductive processes showed that the starvation and feeding protocol periods did not pose a high level of stress to the mites as transcript levels remained comparable between phoretic mites and those completing the protocol. The feeding protocol was used to deliver molecules such as hormone analogs or plasmids. Mites fed with Tebufenozide, an ecdysone analog, had higher transcript levels of shade than untreated or solvent treated mites. In order to extend this feeding protocol, cultured insect cells were incorporated to a final ratio of 1 part cells and 2 parts hemolymph. Although supplementation with Bombyx mori Bm5 cells increased the amount of hemolymph consumed per mite, there was a significant decrease in the percentage of mites that fed and survived. On the other hand, Drosophila melanogaster S2 cells reduced significantly the percentage of mites that fed and survived as well as the amount of hemolymph consumed. The feeding protocol provides a dynamic platform with which to challenge the Varroa mite to establish efficacy of control agents for this devastating honey bee pest.

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Somatic transformation efficiencies and expression patterns using the JcDNV and piggyBac transposon gene vectors in insects

Cited by 17 publications

References 31 publications

“Calling Cards” for DNA-Binding Proteins in Mammalian Cells

“Calling Cards” for DNA-Binding Proteins in Mammalian Cells

Functional characterization of the transcriptional regulatory elements of three highly expressed constitutive genes in the jewel wasp, Nasonia vitripennis

A feeding protocol for delivery of agents to assess development in Varroa mites

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