Generation Of Transgenic Dairy Cattle Using ‘in vitro’ Embryo Production

Krimpenfort, Paul; Rademakers, A; Eyestone, W.H.; A, van der Schans; S, van den Broek; Kooiman, P M; Kootwijk, E.; Platenburg, Gerard; Pieper, Frank; Strijker, R.

doi:10.1038/nbt0991-844

Cited by 205 publications

(122 citation statements)

References 23 publications

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“…Burdon and Wall [30] reported mosaic distribution of injected gene in mouse embryos. In cattle, the mosaicism was also reported in transgenic concepti [7] and bulls [30]. The study with bovine ealry embryos showed 80% of DNA-injected embryos were mosaic [35].…”

Section: Discussionmentioning

confidence: 92%

“…We could not determine whether a fetus with the placenta was transgenic or not. If the embryo was mosaic, the fetus was possible to be non-transgenic, since Krimpenfort et al [7] obtained a mosaic transgenic conceptus, a non-transgenic fetus with a transgenic placenta.…”

Section: Discussionmentioning

confidence: 99%

“…However, since the integration efficiency of injected DNA with host genome is low [4,5], a large number of animals receiving microinjected embryos are required. Application of the microinjection method to produce transgenic cattle has been reported [6][7][8]. The expense of the maintenance for the recipients may defer progress in utilizing transgenic cattle.…”

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confidence: 99%

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Production of Bovine Transgenic Conceptus; Possible Selection of Transgenic Embryos by Polymerase Chain Reaction.

Hoshi

Saeki

Nagao

et al. 1999

Journal of Reproduction and Development

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Abstract. This study was conducted to examine the possibility of selection of transgenic bovine embryos by use of polymerase chain reaction (PCR) following microinjection of exogenous DNA before embryos were transferred to recipient cattle. DNA of bovine α-lactalbumin (α-LA) gene was microinjected into pronuclei of bovine embryos derived from in vitro maturation and fertilization. They were then cultured for 7 to 8 days to the blastocyst stage. To examine persistence of injected DNA during embryo development, embryos were analyzed by PCR at 0, 1, 3, 5 and 7 days following DNA microinjection. To distinguish the injected α-LA from the endogenous gene, gene-gene junction regions of head-to-tail concatemers of the injected DNA, which were formed when linearized DNA were injected into cell nuclei, were amplified by PCR. Injected DNA persisted at high rates (72 to 80%) until 5 days following DNA injection, but the detection rate at the blastocyst stage (20%) was lower than earlier stage (P<0.05), indicating that injected DNA into bovine embryos drastically decreased at the blastocyst stage. To determine if the PCR signals reflect transgenic status, biopsy samples of DNA-injected blastocysts were analyzed. Total of 1,583 embryos were microinjected, and 124 (8.9%) developed to blastocysts. Twelve (11%) out of 109 samples biopsied from microinjected blastocysts were positive by PCR analysis. Eight PCR-positive embryos were transferred to 8 recipients. Also, 31 negative embryos were transferred to recipient heifers. Four out of 8 recipients that received PCR-positive embryos were pregnant and fetuses and placentae were recovered at 60 to 75 days of pregnancy. One of the samples, of which only a placenta was recovered, was found to be transgenic by both PCR analysis and Southern hybridization. The other samples had no injected DNA. On the other hand, 14 fetuses with placentae were recovered from recipients that received PCR-negative embryos. None of these samples were found to carry injected DNA. Our system could not detect single, or head-to-head-or tail-to-tail-joining transgenes; however, the results described here imply selective transfer of potential transgenic embryos to recipient heifers by the PCR selection.

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

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Production of Bovine Transgenic Conceptus; Possible Selection of Transgenic Embryos by Polymerase Chain Reaction.

Hoshi

Saeki

Nagao

et al. 1999

Journal of Reproduction and Development

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“…This technique is used to produce transgenic sheep and pigs (Hammer et al, 1985) and also transgenic cattle (Krimpenfort et al 1991). Using this technique earliest successful creation of transgenic mice was reported in 1980s (Gordon et al, 1980).…”

Section: Pronuclear Microinjectionmentioning

confidence: 99%

Review on current application of genomic biotechnologies to improve livestock and livestock products

2017

Int. J. Adv. Res. Biol. Sci

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An organism's genome is the 'blueprint of life,' the complete set of instructions for making the molecules, cells and tissues that control how an animal can perform throughout its life. Every cell in the body contains the complete genome, encoded in the form of DNA. This review was conducted to give emphasis about the role or the current application of some advance genomic biotechnologies in animal breeding, disease diagnosis and prevention and control strategies. There are opportunities for using molecular genetics to identify genes that are involved in variety of traits. Revolutionary opportunities for the modification of animal performance are being created by the development of new methods (reproductive technology) for embryo manipulation and the application of molecular biology (cloning and transgenesis). Consequently, new approaches are needed to develop improved tools and strategies for prevention and control of infectious diseases in animal agriculture. Among the most effective and successful of these tools are animal vaccines using recombinant deoxyribonucleic acid (rDNA) technologies. In the recent years a profound change has occurred with the introduction of new genomic biotechnological assays for the diagnosis of infectious diseases of livestock and zoonotic pathogens. These new assays include various forms of PCR, genomic sequencing, DNA probes and DNA microarray technology, Nanotechnology, Restriction fragment length polymorphisms and Pulsed field gel electrophoresis. Generally animal genomics is of interest because of its importance to produce high quality food products economically and efficiently to furnish for the increasing supply demand gap all over the world.

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“…Optimisation of the method has been reached by preparing embryos by in vitro maturation of oocytes collected in the slaughterhouse, in vitro fertilization and in vitro development of the embryos after DNA microinjection until the blastocyst stage. This method does not imply the use of embryo donors and it considerably reduces the number of recipients [23].…”

Section: Gene Additionmentioning

confidence: 99%

Transgenesis for the study and the control of lactation

Houdebiné

Rival

Pantano³

et al. 2002

Reprod. Nutr. Dev.

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-The study and the control of milk synthesis are required to decipher the mechanisms of gene expression, to improve milk production, to modify milk composition, to induce a resistance to diseases in the mammary gland and to produce recombinant proteins of pharmaceutical interest. Transgenesis has become a mandatory tool to reach these goals. The use of transgenesis is still limited by the difficulty of adding foreign genes in farm animals and mainly by replacing genes by homologous recombination. Transgene expression is also often ill-controlled. The present paper summarizes the current progress in this field with a particular emphasis on expression vectors for transgenes. lactation / transgenesis / expression vectorsRésumé -La transgénèse pour l'étude et le contrôle de la lactation. L'étude et le contrôle de la synthèse du lait sont nécessaires pour décrypter les mécanismes de l'expression de gènes, pour amé-liorer la production de lait, pour modifier la composition du lait, pour induire une résistance de la glande mammaire contre des maladies infectieuses et pour produire des protéines recombinantes d'intérêt pharmaceutique. La transgénèse est devenue un outil indispensable pour mener à bien ces projets. La mise en oeuvre de la transgénèse est encore limitée par la difficulté d'ajouter des gènes étrangers chez les animaux d'élevage et surtout de remplacer des gènes par recombinaison homologue. L'expression des transgènes est par ailleurs souvent mal contrôlée. Cet article se propose de faire le point dans ce domaine en développant plus particulièrement la mise au point de vecteurs d'expression pour les transgènes. lactation / transgénèse / vecteurs d'expression

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Generation Of Transgenic Dairy Cattle Using ‘in vitro’ Embryo Production

Cited by 205 publications

References 23 publications

Production of Bovine Transgenic Conceptus; Possible Selection of Transgenic Embryos by Polymerase Chain Reaction.

Production of Bovine Transgenic Conceptus; Possible Selection of Transgenic Embryos by Polymerase Chain Reaction.

Review on current application of genomic biotechnologies to improve livestock and livestock products

Transgenesis for the study and the control of lactation

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