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Use of chromatin modifying agents in bovine somatic cell nuclear transfer

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Author(s):
Juliano Rodrigues Sangalli
Total Authors: 1
Document type: Master's Dissertation
Press: Pirassununga.
Institution: Universidade de São Paulo (USP). Faculdade de Zootecnica e Engenharia de Alimentos (FZE/BT)
Defense date:
Examining board members:
Flávio Vieira Meirelles; Mario Binelli; Flávia Lombardi Lopes
Advisor: Flávio Vieira Meirelles
Abstract

Although somatic cell nuclear transfer (SCNT) is a promising tool, its potential use is hampered by the high mortality rates during the development to term of cloned offspring. Abnormal epigenetic reprogramming of donor nuclei after SCNT is thought to be the main cause of this low efficiency. We hypothesized that chromatinmodifying agents (CMAs) targeting chromatin acetylation and DNA methylation could alter the chromatin configuration and turn them more amenable to reprogramming. Thus, bovine fibroblasts were treated with 5-aza-2\'-deoxycytidine (AZA) plus trichostatin (TSA) or hydralazine (HH) plus valproic acid (VPA) whereas, in another trial, cloned bovine zygotes were treated with TSA. The treatment of fibroblasts with either AZA+TSA or HH+VPA increased histone acetylation, but did not affect the level of DNA methylation. However, treatment with HH+VPA decreased cellular viability and proliferation. The use of these cells as nuclear donors showed no positive effect on pre- and post-implantation development. Regarding the treatment of cloned zygotes with TSA, treated one-cell embryos showed an increase in the acetylation patterns, but not in the level of DNA methylation. Moreover, this treatment revealed no positive effect on pre- and post-implantation development. This work provides evidence the treatment of either nuclear donor cells or cloned zygotes with CMAs has no positive effect on pre- and post-implantation development of cloned cattle. (AU)

FAPESP's process: 09/06689-5 - Use of chromatin modification agents in somatic cell nuclear transfer in cattle
Grantee:Juliano Rodrigues Sangalli
Support Opportunities: Scholarships in Brazil - Master