Transformation of frog embryos with a rabbit beta-globin gene.

Rusconi, Sandro; Schaffner, Walter

doi:10.1073/pnas.78.8.5051

Cited by 141 publications

(56 citation statements)

References 42 publications

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“…In this study, the substrate DMA, together with the internal reference plasmid pCml 84, is microinjected into Xenopus laevis stage VI oocytes and fertilized eggs which are known to support homologous recombination and nonhomologous DMA-end joining, respectively (Carroll, 1983;Carroll etal., 1986;Pfeiffer and Vielmetter, 1988; see also Rusconi and Schaffner, 1981). We were able to extend current knowledge about recombination in these cells and show that, under optimal conditions, both events are very fast with the majority of the respective products being produced within the first 20 min after injection.…”

Section: Introductionmentioning

confidence: 69%

“…Fertilized eggs and early embryos of the African clawed frog Xenopus laevis are able to efficiently join any two DNA ends by a nonhomologous recombination mechanism that is referred to as DNA-end joining or end-to-end joining (Pfeiffer and Vielmetter, 1988; see also Rusconi and Schaffner, 1981). From detailed studies with model sub-strates, it is known that this DMA-end joining activity is more sophisticated than originally anticipated.…”

Section: Discussion Homologous Recombination and Nonhomologous Dna-enmentioning

confidence: 98%

“…These colonies are derived from the coinjected circular reference plasmid pCm184. The number of Cm rcolonies during the time course should be a measure for the replication of injected DNA, which has been reported to take place in Xenopus eggs even in the absence of bonafide replication origins (Harland and Laskey, 1980;Rusconi and Schaffner, 1981;Mechali and Kearsey, 1984). The average amplification of -globin gene-containing plasmids was about 50-fold during the long time between zygote and early gastrula (Rusconi and Schaffner, 1981).…”

Section: Very Low Extent Of Dna Replication In Both Oocytes and Fertimentioning

confidence: 99%

“…For oocyte injections typically 1 ng of linearized pReco-σ was used. Fertilized eggs were usually only given 0.2 ng because they do not tolerate higher DMA doses without severe disturbance of proper embryonic development (Rusconi and Schaffner, 1981). Deleterious effects due to the microinjection procedure could be monitored by controlling for proper propagation through the early developmental stages of Xenopus embryogenesis.…”

Section: Fertilized Eggs Almost Exclusively Show Dma-end Joining Whimentioning

confidence: 99%

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Dramatic Changes in the Ratio of Homologous Recombination to Nonhomologous DNA-End Joining in Oocytes and Early Embryos ofXenopus laevis

Hagmann¹,

Adlkofer²,

Pfeiffer³

et al. 1996

Biological Chemistry Hoppe-Seyler

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We have developed a versatile plasmid vector (pReco80% of the homologously recombined product, whereas in eggs a highly efficient DMA-end joining activity predominates Both reactions, homologous recombination and DMA-end joining, are shown to occur quickly, with the majority of the respective products being formed within the first 20 minutes of incubation under optimal conditions. In fertilized eggs, up to 50% of all injected linear DMA molecules are recircularized by DMA-end joining. With high amounts of injected DMA per fertilized egg, DMA-end joining is reduced, presumably due to competition for essential factors, and homologous recombination becomes readily detectable.As there is a sequence of rapid cleavage divisions after fertilization of the egg, the fast and highly efficient DMA-end joining, even though it is error-prone at the junction site, seems to be best suited to cope 1 present address: Institut für Zellbiologie, Swiss Federal Institute of Technology, Zürich-Hönggerberg, CH-8093 Zürich, Switzerland with DMA double-strand breaks that might occur in the genome during early embryogenesis. On the other hand, the long-lived oocytes seem to repair DMA double-strand breaks via homologous recombination. This latter property may be exploited both in Xenopus and in other organisms to achieve homologous integration of exogenous DMA into germ cells for gene targeting.

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Section: Introductionmentioning

confidence: 69%

Section: Discussion Homologous Recombination and Nonhomologous Dna-enmentioning

confidence: 98%

Section: Very Low Extent Of Dna Replication In Both Oocytes and Fertimentioning

confidence: 99%

Section: Fertilized Eggs Almost Exclusively Show Dma-end Joining Whimentioning

confidence: 99%

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Dramatic Changes in the Ratio of Homologous Recombination to Nonhomologous DNA-End Joining in Oocytes and Early Embryos ofXenopus laevis

Hagmann¹,

Adlkofer²,

Pfeiffer³

et al. 1996

Biological Chemistry Hoppe-Seyler

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“…$1 nuclease analysis was performed as described (Weaver and Weissmann, 1979;Rusconi and Schaffner, 1981). Isolation of HCMV IE RNA and Northern blot hybridizations have been described previously (Jahn et al, 1984a).…”

Section: Analysis Of Gene Expressionmentioning

confidence: 99%

A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus

Boshart

Weber

Jahn

et al. 1985

Cell

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1,097

648

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SummaryA strong transcription enhancer was identified in the genomic DNA (235 kb) of human cytomegalovirus (HCMV), a ubiquitous and severe pathogen of the herpesvirus group. Cotransfection of enhancerless SV40 DNA with randomly fragmented HCMV DNA yielded two SV40-HCMV recombinant viruses that had incorporated overlapping segments of HCMV DNA to substitute for the missing SV40 enhancer. Within HCMV, these enhancer sequences are located upstream of the transcription initiation site of the major immediate-early gene, between nucleotides -118 and -524. Deletion studies with the HCMV enhancer, which harbors a variety of repeated sequence motifs, show that different subsets of this enhancer can substitute for the SV40 enhancer. The HCMV enhancer, which seems to have little cell type or species preference, is severalfold more active than the SV40 enhancer. It is the strongest enhancer we have analyzed so far, a property that makes it a useful component of eukaryotic expression vectors.

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Immunolocalization of mitsugumin29 in developing skeletal muscle and effects of the protein expressed in amphibian embryonic cells

et al. 1999

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Transformation of frog embryos with a rabbit beta-globin gene.

Abstract: In order to study the fate and possible expression of foreign DNA Oocytes and eggs of the frog Xenopus laevis have been used for many years as experimental systems for studying the translation of injected mRNAs (reviewed in ref.

Cited by 141 publications

References 42 publications

Dramatic Changes in the Ratio of Homologous Recombination to Nonhomologous DNA-End Joining in Oocytes and Early Embryos ofXenopus laevis

Dramatic Changes in the Ratio of Homologous Recombination to Nonhomologous DNA-End Joining in Oocytes and Early Embryos ofXenopus laevis

A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus

Immunolocalization of mitsugumin29 in developing skeletal muscle and effects of the protein expressed in amphibian embryonic cells

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