Foreign gene expression in activated oocytes and bovine embryos following pAGS-lacZ plasmid microinjection

Gagné, Marc; Pothier, F.; Sirard, Marc-André

doi:10.1016/0093-691x(93)90078-j

Cited by 8 publications

(2 citation statements)

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“…Transcription begins during S-phase of the first cell cycle as demonstrated by microinjection of reporter genes into pronuclei [Ram and Schultz, 1993], expression of integrated transgenes [Matsumoto et al, 1994;Christians et al, 1995], and incorporation of BrUTP into nascent transcripts [Bouniol et al, 1995;Aoki et al, 1997]. Transcription has also been detected at the 1-cell stage in other mammalian species, including the cow [Gagné et al, 1993] and rabbit . This initial zygotic transcription is weak, and results in minor synthesis of a small set of polypeptides that manifest a transient increase restricted to the 2-cell stage [Conover et al, 1991;Latham et al, 1991;Christians et al, 1995;Davis et al, 1996].…”

Section: Activation Of the Zygotic Genome: A Multistep Processmentioning

confidence: 96%

Mouse embryos do not wait for the MBT: Chromatin and RNA polymerase remodeling in genome activation at the onset of development

1998

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Section: Activation Of the Zygotic Genome: A Multistep Processmentioning

confidence: 96%

Mouse embryos do not wait for the MBT: Chromatin and RNA polymerase remodeling in genome activation at the onset of development

1998

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“…An effect of the solution in which the concentrated virus was stored (HEPES-buffered saline containing 10% glycerol) was ruled out by injecting storage solution alone into embryos (data not shown). Moreover, some cytotoxicity has been reported to follow pronuclear injection of DNA, particularly as concentrations are increased [34][35][36]. In human bronchial epithelial cells, the El deletion prevented production of the hexon and fiber proteins, but a subset of cells expressed very high levels of the E2a gene product DBP (DNA-binding protein) [12].…”

Section: Discussionmentioning

confidence: 99%

Adenovirus-Mediated Gene Transfer by Perivitelline Microinjection of Mouse, Rat, and Cow Embryos1

Kubisch

Larson

Eichen

et al. 1997

Biology of Reproduction

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To determine the fate of an episomally expressed transgene, mouse, rat, and cow zygotes were injected into the perivitelline space with approximately 100 pl of buffer containing the replication-defective human adenovirus, AdCMVLacZ/sub360. Viral concentrations ranged from 2.5 to 2.5 x 10(5) plaque-forming units (pfu)/100 pl. As viral titer increased, fewer embryos were able to develop to blastocysts. In the mouse, the percentage of blastocysts formed ranged from 82% in controls to 16% after injection at the highest titer. In the rat and cow, a similar decrease in blastocyst formation was noted (62% to 6% and 26% to 4%, respectively). Reporter gene (galactosidase, LacZ) activity could be detected in mouse embryos after injection at a concentration of only 25 pfu/100 pl, whereas a tenfold higher titer was required in the other two species to observe the blue LacZ reaction product. When examined after 5 (mouse), 6 (rat), or 9 (cow) days of in vitro culture, the proportion of LacZ-positive embryos ranged from 15% to 96%, 6% to 76%, and 18% to 58% in mouse, rat, and cow embryos, respectively, depending upon viral concentration. However, a large percentage of positive embryos proved to be expression mosaics, the degree of which was likewise dependent on titer. While none of the embryos showed LacZ activity at 30 h after injection, 70% of mouse, 8% of rat, and 20% of cow embryos expressed the reporter gene at 42 h. Delaying the timing of injection revealed that the efficiency with which mouse and rat embryos could be infected decreased with increasing degree of differentiation. Only 35% and 18% of mouse embryos expressed the reporter gene after injection at the morula or blastocyst stage, respectively. A similar drop in efficiency was noted in rat embryos when injections took place at the 8-cell, morula, or blastocyst stage, with 70%, 33%, and 9% of embryos, respectively, subsequently showing LacZ activity. Likewise, advanced development resulted in a decrease in the efficiency of viral-mediated gene transfer in cow embryos, with 100%, 78%, and 68% of embryos being positive after injection at the 8-cell, morula, or blastocyst stage, respectively. These results demonstrate that a human adenovirus can be used to express a reporter gene transiently in nonhuman embryos.

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Expression of a Foreign Gene Introduced Into Bovine Embryos at Early Stages of Development

Muramatsu

Nakamura

Okumura

1994

Animal Cell Technology: Basic &Amp; Applied Aspects

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Foreign gene expression in activated oocytes and bovine embryos following pAGS-lacZ plasmid microinjection

Cited by 8 publications

References 0 publications

Mouse embryos do not wait for the MBT: Chromatin and RNA polymerase remodeling in genome activation at the onset of development

Mouse embryos do not wait for the MBT: Chromatin and RNA polymerase remodeling in genome activation at the onset of development

Adenovirus-Mediated Gene Transfer by Perivitelline Microinjection of Mouse, Rat, and Cow Embryos1

Expression of a Foreign Gene Introduced Into Bovine Embryos at Early Stages of Development

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