Abstract
Objective: To evaluate the effect of treatment with trichostatin-A (TSA) on the production of bovine embryos, expressing the gene of the green fluorescent protein (GFP) generated by SCNT. Materials: 164 oocytes were distributed in three treatments, NT-GFP: newly reconstructed zygotes with genetically modified cells and not subject to TSA. NTTrico-GFP: newly reconstructed zygotes with genetically modified cells and subjected to TSA. PART: Zygotes generated by parthenogenetic activation, used as a control for the process of oocyte activation and culture of embryos. The rates of cleavage, blastocysts, and embryos that expressed GFP were assessed by contingency tables and chi-square tests. Results: The percentage of cleavage in the zygotes in the NT-GFP treatment was greater but did not vary significantly from the NT-Trico-GFP treatment. However, this last treatment had a higher percentage of blastocyst formation (p=0.077). The percentage of blastocysts from cleaved zygotes, the produced embryos were significantly higher (p<0.05) for the NT-Trico-GFP treatment than for the NT-GFP. In both treatments, all the blastocysts generated expressed the GFP protein. Conclusions: TSA improves the embryonic development of clones of genetically modified cattle that express GFP. Keywords: Embryonic Development, Epigenetic Modification, Nuclear Transfer
Highlights
Among the different techniques of genetic manipulation, there are several methods that can be used to generate transgenic animals
For the production of embryos by SCNT, the fibroblasts genetically modified with green fluorescent protein (GFP) for nuclear transfer (NT)-GFP and NT-Trico-GFP treatments were thawed and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% fetal bovine serum (FBS) for 1 day
The results of the viral translation in cells depend on several factors, such as initial cell concentration, type of cell; viral concentration added to the culture, among others, the positivity of the cells that express the transgene after translation is variable[12]
Summary
Among the different techniques of genetic manipulation, there are several methods that can be used to generate transgenic animals Among these we have, the transfer by means of viral vectors, DNA transfer mediated by spermatozoa, a nuclear transfer from transfected somatic cells (SCNT)[1] and CRISPR/Cas[92] technology. In the phase of cellular reprogramming, cell division must begin and it has as limitation the chromosomal anomalies that can occur[11,12,13,14] and that lead to the death of cloned animals due to the epigenetic alterations that affect the structure, composition and remodeling of the chromatin that defines and maintains the accessibility and transcription of the genetic information contained in the DNA10. Reprogramming involves changes in nuclear material, such as the structure of chromatin and re-methylation of DNA These events are necessary for the expression of the genome, silencing genes from the old differentiated cell and stimulating genes characteristic of embryonic cells[7]. The objective of this research was to evaluate the effect of treatment with TSA on the production of bovine embryos, expressing the green fluorescent protein (GFP) gene generated by SCNT
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call