Germline transgenesis of zebrafish using the medaka <i>Tol1</i> transposon system

Koga, Akihiko; Cheah, Foong Koon; Hamaguchi, Satoshi; Yeo, Gare Hoon; Chong, Samuel S.

doi:10.1002/dvdy.21688

Cited by 47 publications

(39 citation statements)

References 23 publications

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“…1. The two elements are independent transposase-substrate systems, however: the Tol1 transposase does not catalyze transposition of Tol2, and the Tol2 transposase does not serve as an enzyme for Tol1 (Koga et al, 2008). If they were not independent of each other, the conspicuous difference in their evolutionary stages observed in the present study and our previous studies would not have been generated.…”

Section: Discussioncontrasting

confidence: 47%

“…Detection of the latter requires a complex approach because no one can anticipate where the element will land. In the present work, we only examined excision, because the Tol1 transposase has already been demonstrated to catalyze both excision and insertion (Koga et al, 2008).…”

Section: Excision Assay With Culture Cellsmentioning

confidence: 99%

“…pDon[Tol1] is a plasmid used in our previous study (Koga et al, 2008). It carries the terminal regions of Tol1 with a green fluorescent protein (GFP) gene reporter cassette between them (Fig.…”

Section: Excision Assay With Culture Cellsmentioning

confidence: 99%

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Distribution of complete and defective copies of the Tol1 transposable element in natural populations of the medaka fish Oryzias latipes

et al. 2009

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DNA-based transposable elements are present in the genomes of various organisms, and generally occur in autonomous and nonautonomous forms, with a good correspondence to complete and defective copies, respectively. In vertebrates, however, the vast majority of DNA-based elements occur only in the nonautonomous form. Until now, the only clear exception known has been the Tol2 element of the medaka fish, which still causes mutations in genes of the host species. Here, we report another exception: the Tol1 element of the same species. This element was thought likely to be a "dead" element like the vast majority of vertebrate elements, but recent identification of an autonomous Tol1 copy in a laboratory medaka strain gave rise to the possibility that the element is still "alive" in medaka natural populations. We examined variation in the structure of Tol1 copies through genomic Southern blot analysis, and revealed that 10 of the 32 fish samples examined contained full-length Tol1 copies in their genomes. The frequency at which these copies occur among Tol1 copies is at most 0.5%, yet some of them still have the ability to produce a functional transposase. The medaka fish thus harbors two active DNA-based elements in its genome, and is in this respect unique among vertebrates.

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Section: Discussioncontrasting

confidence: 47%

Section: Excision Assay With Culture Cellsmentioning

confidence: 99%

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Distribution of complete and defective copies of the Tol1 transposable element in natural populations of the medaka fish Oryzias latipes

et al. 2009

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“…5A). In this case, we utilized transiently injected Tol1 transposons (Koga et al, 2008) to avoid mobilizing the Tg(cdh5:gal4ff ) mu101 transgene, which is flanked by Tol2 direct repeat elements (Bussmann et al, 2011). flt4 C562Δ ;Tg (cdh5:gal4ff ) mu101 mutant embryos injected with a control Tol1 transgene did not form lympho-venous sprouts and failed to induce Prox1 expression in the PCV while displaying robust endothelialexpressed nlstdTomato (Fig.…”

Section: Erk Activation Is Necessary and Sufficient To Induce Differementioning

confidence: 99%

“…A multisite Gateway destination cassette (attR4-ccdb-cam r -attR3) followed by a polyadenylation sequence and flanked by Tol1 direct repeats (Koga et al, 2008) was synthesized in pUC57amp (Genewiz) and is referred to as pToneDest. A cDNA encoding full-length Tol1 transposase (Koga et al, 2008) and a polyadenylation sequence was synthesized in pUC57 downstream of a T7 promoter and is referred to as pToneTpase.…”

Section: Plasmid Constructionmentioning

confidence: 99%

Vegfc acts through ERK to induce sprouting and differentiation of trunk lymphatic progenitors

et al. 2016

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There was an error published in Development 143, 3785-3795.In the Materials and Methods section we provided an incorrect serine residue number for the map2k2b mutation. The corrected section of text is as follows:An activated myc-tagged form of MEK was generated from zebrafish map2k2b by mutating serines 221 and 225 to aspartic acid (Schramek et al., 1997) by PCR, followed by BP cloning to generate pME-myc-act-map2k2b.In addition, we have since verified that the pME-myc-act-map2k2b plasmid used in the original studies and submitted to Addgene contains the correct sequence. This plasmid contains several nucleotide changes in comparison to the GenBank reference sequence (accession number: NM_001128281). However, all of these changes have been noted in a previous annotation of single nucleotide polymorphisms in the zebrafish and are naturally occurring variants (Butler et al., 2015).We apologize for any confusion that this may have caused and we thank the researchers who discovered the error for bringing it to our attention. ABSTRACTVascular endothelial growth factor C (Vegfc) activates its receptor, Flt4, to induce lymphatic development. However, the signals that act downstream of Flt4 in this context in vivo remain unclear. To understand Flt4 signaling better, we generated zebrafish bearing a deletion in the Flt4 cytoplasmic domain that eliminates tyrosines Y1226 and 1227. Embryos bearing this deletion failed to initiate sprouting or differentiation of trunk lymphatic vessels and did not form a thoracic duct. Deletion of Y1226/7 prevented ERK phosphorylation in lymphatic progenitors, and ERK inhibition blocked trunk lymphatic sprouting and differentiation. Conversely, endothelial autonomous ERK activation rescued lymphatic sprouting and differentiation in flt4 mutants. Interestingly, embryos bearing the Y1226/7 deletion formed a functional facial lymphatic network enabling them to develop normally to adulthood. By contrast, flt4 null larvae displayed hypoplastic facial lymphatics and severe lymphedema. Thus, facial lymphatic vessels appear to be the first functional lymphatic network in the zebrafish, whereas the thoracic duct is initially dispensable for lymphatic function. Moreover, distinct signaling pathways downstream of Flt4 govern lymphatic morphogenesis and differentiation in different anatomical locations.

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Site‐directed zebrafish transgenesis into single landing sites with the phiC31 integrase system

Mosimann

Puller

Lawson

et al. 2013

Developmental Dynamics

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Background Linear DNA-based and Tol2-mediated transgenesis are powerful tools for the generation of transgenic zebrafish. However, the integration of multiple copies or transgenes at random genomic locations complicates comparative transgene analysis and makes long-term transgene stability unpredictable with variable expression. Targeted, site-directed transgene integration into pre-determined genomic loci can circumvent these issues. The phiC31 integrase catalyzes the unidirectional recombination reaction between heterotypic attP and attB sites and is an efficient platform for site-directed transgenesis. Results We report the implementation of the phiC31 integrase-mediated attP/attB recombination for site-directed zebrafish transgenics of attB-containing transgene vectors into single genomic attP landing sites. We generated Tol2-based single-insertion attP transgenic lines and established their performance in phiC31 integrase-catalyzed integration of an attB-containing transgene vector. We found stable germline transmission into the next generation of an attB reporter transgene in 34% of all tested animals. We further characterized two functional attP landing site lines and determined their genomic location. Our experiments also demonstrate tissue-specific transgene applications as well as long-term stability of phiC31-mediated transgenes. Conclusions Our results establish phiC31 integrase-controlled site-directed transgenesis into single, genomic attP sites as space-, time-, and labor-efficient zebrafish transgenesis technique. The described reagents are available for distribution to the zebrafish community.

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Germline transgenesis of zebrafish using the medaka Tol1 transposon system

Cited by 47 publications

References 23 publications

Distribution of complete and defective copies of the Tol1 transposable element in natural populations of the medaka fish Oryzias latipes

Distribution of complete and defective copies of the Tol1 transposable element in natural populations of the medaka fish Oryzias latipes

Vegfc acts through ERK to induce sprouting and differentiation of trunk lymphatic progenitors

Site‐directed zebrafish transgenesis into single landing sites with the phiC31 integrase system

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