Production of dairy goat embryos, by nuclear transfer, transgenic for human acid β-glucosidase

Zhang, Y.L.; Wan, Yongjie; Wang, Z.Y.; Xu, Dachao; Pang, Xinzhi; Li, Meng; Wang, L.H.; Zhong, Bushuai; Wang, F.

doi:10.1016/j.theriogenology.2009.11.008

Cited by 29 publications

(14 citation statements)

References 38 publications

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“…After a few days, the structures could no longer be observed. These findings corroborate with Baguisi et al (1999) and Zhang et al (2010). According to Baguisi et al (1999), more than 2/3 of their clone pregnancies were not viable, and such "embryonic structures" were observed in the uterus until day 55 of gestation.…”

Section: Discussionsupporting

confidence: 87%

Effects of oocyte source, cell origin, and embryo reconstruction procedures on in vitro and in vivo embryo survival after goat cloning

Feltrin¹,

Aguiar²,

Calderón³

et al. 2017

Anim Reprod

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The birth of cloned goats has been well documented, but the overall goat cloning efficiency by somatic cell nuclear transfer procedures is still low, which may be further intensified in extreme environments. The aim of this study was to produce cloned goats under the conditions of the Brazilian SemiArid region, in a transgenic program for the expression of human lysozyme in the milk to target childhood diarrhea and malnutrition, comparing the effects of oocyte source, cell type, and embryo reconstruction procedures on in vitro and in vivo embryo survival after cloning by micromanipulation or by handmade cloning. The use of in vitro-matured oocytes resulted in more viable embryos after cloning than in vivo-matured cytoplasts, but no differences in pregnancy rates on day 23 were seen between oocyte sources (77.5 vs. 77.8%, respectively). The presence or absence of the zona pellucida for embryo reconstruction (78.8 vs. 76.0%, respectively) did not affect pregnancy outcome after transfer. However, pregnancy rate on day 23 was higher for embryos chemically activated by a conventional than a modified protocol (88.1 vs. 50.0%), and for embryos reconstructed with mesenchymal stem cells and fetal fibroblasts (100.0 and 93.3%) than with adult fibroblasts (64.7%). Although most pregnancies were lost, the birth of a cloned female was obtained from embryos reconstructed by micromanipulation using non-transgenic control cells and in vitro-matured oocytes with intact zona pellucida, after conventional activation and transfer at the 1-cell stage.

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Section: Discussionsupporting

confidence: 87%

Effects of oocyte source, cell origin, and embryo reconstruction procedures on in vitro and in vivo embryo survival after goat cloning

Feltrin¹,

Aguiar²,

Calderón³

et al. 2017

Anim Reprod

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“…However, because we observed embryos at days 3 and 8 postreconstruction to detect cleavage and blastocyst rates, a direct comparison between the percentages of the mixed population of morula and blastocysts and merely blastocysts is not possible. Of note, although the latter group (Zhang et al, 2010) transferred transgenic embryos at the early stages to 16 recipients, pregnancy was only confirmed at day 40 in two goats and did not confirm later. Higher NT blastocyst development in this study may have been a result of systematic investigations and optimizations performed to provide the goat-specific procedures of IVM, SCNT, and IVC.…”

Section: Scnt Efficiencymentioning

confidence: 85%

“…In terms of blastocyst development, the results of this study (21-33%) are higher than studies that have cultured NT embryos either in vivo (Behboodi et al, 2004: 20% morulae and blastocyst;Zou et al, 2001: 7.8% blastocyst) or in vitro (Behboodi et al, 2004: 15% morulae and blastocyst;Chen et al, 2007: 8.8% blastocyst;Lan et al, 2006: 17.5% blastocyst;Ohkoshi et al, 2003: 3% blastocyst;Zhang et al, 2004: 6.8% blastocyst). In a recent study, Zhang et al (2010) reported 36.4% versus 38.9% morulae and blastocyst from NT embryos derived from transgenic fetal fibroblasts versus nontransfected control cells. However, because we observed embryos at days 3 and 8 postreconstruction to detect cleavage and blastocyst rates, a direct comparison between the percentages of the mixed population of morula and blastocysts and merely blastocysts is not possible.…”

Section: Scnt Efficiencymentioning

confidence: 98%

Development of an Optimized Zona-Free Method of Somatic Cell Nuclear Transfer in the Goat

Nasr-Esfahani

Hosseini

Hajian

et al. 2011

Cellular Reprogramming

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The purpose of this study was to develop an improved zona-free method of goat somatic cell nuclear transfer (SCNT) that has both ease of operation and efficiency. The main steps involved were: (1) optimization of in vitro oocyte maturation, (2) parthenogenetic activation of zona-free oocytes, (3) SCNT of zona-free anaphase II-telophase II (AII-TII) oocytes that subverted the need for long term UV-exposure of the oocytes, and (4) in vitro culture of groups of cloned embryos in wells in a highly efficient continuous serum-free embryo medium to the blastocyst stage before transfer to the recipients. Percentages of transgenic blastocyst production were 22.3 and 33.1% for adult and fetal cell lines, respectively. After transfer of cloned and transgenic blastocysts, 28.6 and 36.4% of the recipients were confirmed pregnant and 75 and 33.3% of the pregnancies resulted in the delivery of viable offspring, respectively. To our knowledge, this is the first report of successful live and survived birth of cloned and transgenic offspring through a whole procedure of in vitro oocyte maturation and embryo development to the blastocyst stage, and in this study the in vitro efficiencies of cloned and transgenic embryo production were higher than the available reports.

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“…Oocyte collection and in vitro maturation, SCNT, activation of reconstructed embryos, culture of cloned embryos, embryo transfer, and pregnancy diagnosis were performed as described previously (Lazaris et al, 2006;Zhang et al, 2010).…”

Section: Production Of Transgenic Dairy Goats By Scntmentioning

confidence: 99%

Identification of transgenic cloned dairy goats harboring human lactoferrin and methylation status of the imprinted gene IGF2R in their lungs

Zhang¹,

Zhang²,

Wan³

et al. 2015

Genet. Mol. Res.

Self Cite

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ABSTRACT. Dairy goat is a good model for production of transgenic proteins in milk using somatic cell nuclear transfer (SCNT). However, animals produced from SCNT are often associated with lung deficiencies. We recently produced six transgenic cloned dairy goats harboring the human lactoferrin gene, including three live transgenic clones and three deceased transgenic clones that died from respiratory failure during the perinatal period. Imprinted genes are important regulators of lung growth, and may be subjected to faulty reprogramming. In the present study, first, microsatellite analysis, PCR, and DNA sequence identification were conducted to confirm that these three dead kids were genetically identical to the transgenic donor cells. Second, the CpG island methylation profile of the imprinted insulinlike growth factor receptor (IGF2R) gene was assessed in the lungs of the three dead transgenic kids and the normally produced kids using bisulfite sequencing PCR. In addition, the relative mRNA level of Y.L. Zhang et al. 11100©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (3): 11099-11108 (2015) IGF2R was also determined by real-time PCR. Results showed that the IGF2R gene in the lungs of the dead cloned kids showed abnormal hypermethylation and higher mRNA expression levels than the control, indicating that aberrant DNA methylation reprogramming is one of the important factors in the death of transgenic cloned animals.

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Production of dairy goat embryos, by nuclear transfer, transgenic for human acid β-glucosidase

Cited by 29 publications

References 38 publications

Effects of oocyte source, cell origin, and embryo reconstruction procedures on in vitro and in vivo embryo survival after goat cloning

Effects of oocyte source, cell origin, and embryo reconstruction procedures on in vitro and in vivo embryo survival after goat cloning

Development of an Optimized Zona-Free Method of Somatic Cell Nuclear Transfer in the Goat

Identification of transgenic cloned dairy goats harboring human lactoferrin and methylation status of the imprinted gene IGF2R in their lungs

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