Treating donor cells with 2-PCPA corrects aberrant histone H3K4 dimethylation and improves cloned goat embryo development

Abstract

SCNT: somatic cell nuclear transfer; H3K4me2: H3K4 dimethylation; 2-PCPA: trans-2-Phenylcyclopropylamine; LSD1: lysine-specific demethylase 1; GFFs: goat fetal fibroblast cells; IVF: in vitro fertilization; iPS: induced pluripotent stem; PBS: phosphate-buffered saline; IVM: in vitro maturation; RNAPII: RNA polymerase II; HMTs: histone methyltransferase.

Effects of Crotonylation on Reprogramming of Cashmere Goat Somatic Cells with Different Differentiation Degrees

“…This result also demonstrated that, unlike the process in IVF embryos, epigenetic reprogramming in somatic cells is incomplete, leading to abnormal development in SCNT embryos. The researchers attempted to implement some repair means to correct the reprogramming of somatic cells and the aberrant development of cloned embryos [ 39 , 40 ]. Previous studies have found that somatic cell reprogramming and the developmental competence of cloned embryos were improved by correcting epigenetic modifications [ 41 ].…”

Generating Cloned Goats by Somatic Cell Nuclear Transfer—Molecular Determinants and Application to Transgenics and Biomedicine

“…Those that have been tested include cells stemming from several types of tissues collected from both caprine fetuses and adult animals of both sexes and of different ages. Among the nuclear donor cells (NDCs) used for SCNT procedures, mention should be made of: (1) in vitro cultured (transgenic or nontransgenic) fetal dermal fibroblasts [ 12 , 16 , 17 , 18 , 19 , 20 , 21 , 22 ]; (2) juvenile and adult dermal fibroblasts [ 14 , 23 , 24 , 25 , 26 ]; (3) mural granulosa cells isolated from antral ovarian follicles [ 18 , 27 ]; (4) cumulus oophorus cells [ 18 , 27 , 28 ].…”

“…One of the strategies to reverse advanced alterations in the pattern of epigenetic covalent modifications within somatic cell nuclei, which encompass rapid DNA methylation and a decrease in the quantitative profile of histone protein acetylation, appears to be the exposure of NDCs, SCNT-derived oocytes, and corresponding embryos to reversible agents inhibiting biocatalytic activity of DNA methyltransferases (DNMTs) and/or histone deacetylases (HDACs). The use of nonselective or selective promoters of epigenetically determined transcriptional activity of genomic DNA in both in vitro cultured NDCs and cloned embryos is supposed to be an approach that allows for proper reprogramming of somatic cell nuclei [ 7 , 12 , 14 ]. Exogenously modulating the epigenetic memory profile of genomic DNA appears to contribute to successfully reversing the “transcriptional clock” of a differentiated somatic cell nucleus to the status of a cell nucleus characteristic of a totipotent or pluripotent embryonic cell.…”

“…The aforementioned strategies may considerably modify the epigenetically determined reprogramming of the somatic cell nuclear genome in SCNT-derived embryos. The final results of these innovative solutions turn out to be significant enhancements of the pre- and/or postimplantation developmental competence and an improvement in the molecular quality of cloned embryos in mammals, including the domestic goat [ 12 , 13 , 14 , 42 , 43 , 44 ].…”

“…This calls for studies aimed at the precise determination of the conditions that facilitate epigenetic reprogramming in the nuclear donor cell genome during the pre- and postimplantation development of SCNT-generated embryos and fetuses of different mammalian species, including the domestic goat [ 5 , 6 , 7 , 8 , 9 , 10 ]. Promising results were achieved by investigations that focused on the use of extrinsic nonselective agents for stimulating the epigenetically regulated transcriptional activity of genomic DNA in both nuclear donor somatic cells and SCNT-cloned embryos [ 11 , 12 , 13 , 14 , 15 ].…”