+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

Generation of transgenic dairy cattle from transgene-analyzed and sexed embryos produced in vitro

Generation of transgenic dairy cattle from transgene-analyzed and sexed embryos produced in vitro

Bio/Technology 12(6): 606-608

We have generated a transgenic calf from in vitro produced bovine embryos which had undergone transgene analysis and sexing prior to the embryo transfer. Bovine oocytes were isolated from slaughter-house-derived ovaries, matured and fertilized in vitro and subsequently microinjected with a dam-methylated gene construct consisting of genomic sequences encoding human erythropoietin and governed by bovine alpha S1-casein regulatory sequences. After 6 to 7 days in culture, the embryos were biopsied and while the embryo remained in culture, the biopsy was subjected to transgene analysis and sexing. The transgene analysis was accomplished with a combined treatment of the embryo lysates with DpnI restriction endonuclease and Bal31 exonuclease followed by polymerase chain reaction (PCR). The transgene analysis was based on the fact that DpnI only cleaves its recognition sequence if the adenine in the sequence is methylated. Pregnancy was induced by the transfer of three viable female embryos with a distinct transgene signal to a hormonally synchronized heifer recipient. Amniotic fluid analysis performed two months after the embryo transfer confirmed the presence of the transgene. The calf born was found to be transgenic by PCR analysis from blood, ear and fetal membranes. The presence of the transgene was also confirmed by Southern blotting.

Accession: 043186789

Download citation: RISBibTeXText

PMID: 7764950

DOI: 10.1038/nbt0694-606

Related references

Transgenic cattle from transgene-analyzed and sexed embryos. Biotecnologia Aplicada 13(1): 33, 1996

Generation of transgenic cattle from microinjected embryos produced in vitro. 1994

Blastocyst viability and generation of transgenic cattle following freezing of in vitro produced, DNA-injected embryos. Animal Reproduction Science 63(1/2): 53-63, 2000

Use of sexed sorted semen for fixed-time artificial insemination or fixed-time embryo transfer of in vitro-produced embryos in cattle. Theriogenology 86(3): 888-893, 2017

Generation of transgenic dairy cattle using 'in vitro' embryo production. Bio/Technology 9(9): 844-847, 1991

Effect of concentration of sexed bovine sperm sorted at 40 and 50 psi on developmental capacity of in vitro produced embryos. Theriogenology 59(1): 506, January 1, 2003

Birth of double-muscled Belgian Blue calves after transfer of in vitro produced embryos into dairy cattle. Theriogenology 41(4): 855-867, 1994

Molecular characterization of transgene integration by next-generation sequencing in transgenic cattle. Plos One 7(11): E50348, 2013

Efficacy of timed embryo transfer with fresh and frozen in vitro produced embryos to increase pregnancy rates in heat-stressed dairy cattle. Journal of Dairy Science 82(11): 2369-2376, 1999

Selection of in vitro produced, transgenic embryos by nested PCR for efficient production of transgenic goats. Theriogenology 56(4): 545-556, 2001

Effect of transfer of one or two in vitro-produced embryos and post-transfer administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle. Theriogenology 66(2): 224-233, 2005

Morphological and Gene Expression Changes in Cattle Embryos from Hatched Blastocyst to Early Gastrulation Stages after Transfer of In Vitro Produced Embryos. Plos One 10(6): E0129787, 2016

Effects of trichostatin A on in vitro development and transgene function in somatic cell nuclear transfer embryos derived from transgenic Clawn miniature pig cells. Animal Science Journal 81(5): 558-563, 2011

Normality of calves produced from IVF with sexed sperm and culturing embryos with insulin. Theriogenology 57(1): 754, January 2, 2002

Trimethylation of histone 3 at lysine 4 in cryopreserved bovine embryos produced in vivo with sexed semen. Theriogenology 86(8): 1944-1952, 2017