Persistence and replication of plasmid DNA microinjected into early embryos of Xenopus laevis

Marini, Nicholas J.; Etkin, Laurence D.; Benbow, Robert M.

doi:10.1016/0012-1606(88)90328-4

Cited by 51 publications

(48 citation statements)

References 48 publications

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“…These included not only supercoiling and linearization of the circular molecules but also the formation of both covalently (concatemers) or non-covalently (concatenates) linked multimers. Similar observations have been made in higher vertebrates after transfer of circular plasmid DNA into cells in vitro or in vivo (Folgeret al 1982;Brinster et al 1985;Marini et al 1988) and in previous studies in fish embryos (Chong and Vielkind 1989). However, in sea urchins it has been shown that injected circular plasmid DNA is neither concatemerized nor replicated .…”

Section: Discussionsupporting

confidence: 81%

“…In Xenopus and mammalian cells, on the other band, the fonnation of linear, exclusively head-to-tail multimers after injection of circular DNA has been previously shown (Rusconi and Schaffner 1981;Folger et al 1982;Etkin et al 1984). A homologaus recombination mechanism was postulated that, in the case of Xenopus, is also thought tobe responsible for the formation ofhead-to-tail concatemers after injection of linearized plasmid DNA (Marini et al 1988). In our studies using Southern blot analysis we could not detect formation of head-to-head or tail-to-tail concatemers because digestion of D~A extracted from injected embryos with an enzyme that cuts the introduced plasmid once showed no additional bands (Fig.…”

Section: Discussionmentioning

confidence: 93%

“…Replication was independent of specific sequences, but dependent on number and size of injected plasmid molecules. However, replication of foreign DNA was found only up to gastrulation and occurred mainly in hmw concatemers (Rusconi and Schaffner 1981 ;Etkin et al 1984;Marini et al 1988). In older embryos only limited amounts of foreign DNA persisted due to degradation.…”

Section: Discussionmentioning

confidence: 99%

“…Such processes could be responsible (i) for a mosaic distribution of foreign DNA to daughter cells, and (ii) for replication and transcription of the foreign DNA by host nuclear enzymes. Additionally, it has been suggested that maternal enzymes are responsible for the conformational changes in and replication of the foreign DNA in the very early embryo (Zierler et al 1985;Marini et al 1988). This might not account for the findings in the medaka system, because here replication occurs also in later stages.…”

Section: Discussionmentioning

confidence: 99%

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Transient expression of foreign DNA during embryonic and larval development of the medaka fish (Oryzias latipes)

Winkler

Vielkind²,

Schartl

1991

Mol Gen Genet

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Summary. Species of small fish are becoming useful tools for studies on vertebrate development. Wehave investigated the developing embryo of the Japanese medaka for its application as a transient expression system for the in vivo analysis of gene regulation and function. The temporaland spatial expression patterns ofbacterial chloramphenicol acetyltransferase and galactosidase reporter genes injected in supercoiled plasmid form into the cytoplasm of one cell of the two-cell stage embryo was promoter-specific. The transient expression was found to be mosaic within the tissue and organs reflecting the unequal distribution of extrachromosomal foreign DNA and the intensive cell mixing movements that occur in fish embryogenesis. The expression data are consistent with data on DNA fate. Foreign DNA persisted during embryogenesis and was still detectable in some 3-and 9-month-old adult fish; it was found in high molecular weight form as weil as in circular plasmid conformations. The DNA was replicated during early and late embryogenesis. Our data indicate that the developing medaka embryo is a powerful in vivo assay system for studies of gene regulation and function.

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Section: Discussionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Transient expression of foreign DNA during embryonic and larval development of the medaka fish (Oryzias latipes)

Winkler

Vielkind²,

Schartl

1991

Mol Gen Genet

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“…We hypothesize that after Ds excision in the cytoplasm, the external vector DNA undergoes fairly accurate end-joining (presumably followed by amplification), resulting in generation of predominantly uniform products. The presence of DNA ligases in the cytoplasm and the concatenation and amplification of microinjected linear exogenous DNA were previously reported (Soderhall and Lindahl 1975;Prigent et al 1987;Marini et al 1988). In the nucleus, the vector DNA from which the Ds excised may be processed differently (e.g., distinct repair mechanism, nuclease degradation).…”

Section: Discussionmentioning

confidence: 81%

Trans-Kingdom Transposition of the Maize Dissociation Element

et al. 2006

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Transposons are very valuable tools for genetic manipulation. However, the number of transposable elements that have been suitably adapted for experimental use is insufficient and the spectrum of heterologous hosts in which they have been deployed is restricted. To date, only transposons from animal hosts have been utilized in heterologous animal species and transposons of plant origin have been used in plant genetics. There has been no experimental evidence that any of the known elements could transpose in hosts belonging to both kingdoms. Here we demonstrate that the maize Dissociation (Ds) element is capable of effective Activator (Ac) transposase-mediated transposition in the zebrafish Danio rerio, yielding remarkable germline transmission rates. In addition, mammalian cells were also found to be conducive to Ds transposition. Furthermore, we demonstrate that nuclear localization of Ac transposase is essential for genomic Ds transposition. Our results support the hypothesis that Ac/Ds elements do not rely on hostspecific factors for transposition and that host factors involved in their mobility mechanism are widely conserved. Finally, even in vertebrate cells, the Ac/Ds system displays accurate transposition, largefragment carrying capacity, high transposition frequencies, efficient germline transmission, and reporter gene expression, all of which are advantageous for various genetic applications and animal biotechnology.

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On the nature of origins of DNA replication in eukaryotes

1992

Self Cite

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Chromosomal origins of DNA replication in higher eukaryotes differ significantly from those of E. coli (oriC) and the tumor virus, SV40 (ori sequence). Initiation events appear to occur throughout broad zones rather than at specific origin sequences. Analysis of four chromosomal origin regions reveals that they share common modular sequence elements. These include DNA unwinding elements, pyrimidine tracts that may serve as strong DNA polymerase-primase start sites, scaffold associated regions, transcriptional regulatory sequences, and, possibly, initiator protein binding sites and inherently destabilized regions. Based on the novel organization of chromosomal origin regions, we propose a model for initiation of DNA replication in higher eukaryotes. Unwinding of duplex DNA during initiation may be uncoupled, both temporally and spatially, from DNA synthesis, resulting in transient single-stranded intermediates that function in lieu of conventional replication forks during chromosomal DNA replication. DNA synthesis begins subsequently at multiple sites within the unwound regions rather than at specific origin sequences.

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Persistence and replication of plasmid DNA microinjected into early embryos of Xenopus laevis

Cited by 51 publications

References 48 publications

Transient expression of foreign DNA during embryonic and larval development of the medaka fish (Oryzias latipes)

Transient expression of foreign DNA during embryonic and larval development of the medaka fish (Oryzias latipes)

Trans-Kingdom Transposition of the Maize Dissociation Element

On the nature of origins of DNA replication in eukaryotes

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