Enhancement of Chromatin and Epigenetic Reprogramming in Porcine SCNT Embryos—Progresses and Perspectives

Glanzner, Werner Giehl; Macedo, Mariana Priotto de; Gutierrez, Karina; Bordignon, Vilceu

doi:10.3389/fcell.2022.940197

Cited by 7 publications

(5 citation statements)

References 150 publications

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“…These oocytes may have been responsible for the high rate of apoptosis when they reach the blastocyst. The low developmental capacity of somatic cell nuclear transfer has been primarily attributed to incomplete reprogramming of the donor ( Zhang et al, 2020 ; Glanzner et al, 2022 ). Donor cells injected for the production of CITP-derived tetraploid embryos also have processing similar to somatic cell nuclear transfer.…”

Section: Discussionmentioning

confidence: 99%

Tetraploid embryo aggregation produces high-quality blastocysts with an increased trophectoderm in pigs

Lee,

Cai,

Kim

et al. 2023

Front. Cell Dev. Biol.

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Tetraploid complementation is an ideal method for demonstrating the differentiation potential of pluripotent stem cells. In this study, we selected the most efficient tetraploid production method for porcine embryos and investigated whether tetraploid blastomere aggregation could enhance the quality of tetraploid embryos. Three methods were investigated to produce tetraploid embryos: First, tetraploid embryos were produced using electro-fusion of two-cell stage parthenogenetic blastomere (FUTP). Second, somatic cell was injected into the mature oocyte and fused to produce tetraploid embryos. Third, oocytes were matured with Cytochalasin B (CB) for the late 22 h of in vitro maturation to inhibit the first polar body (PB1). Following that, non-PB1 oocytes were treated with CB for 4 h after parthenogenetic activation. There was no significant difference in the blastocyst development rate and tetraploid production rate of the embryos produced through the three methods. However, FUTP-derived blastocysts had a significantly lower percentage of apoptotic cells compared to other methods. The developmental competence of embryos, expression of trophectoderm cell marker genes, and distribution of YAP1 protein were investigated in tetraploid embryos produced using the FUTP method. The FUTP method most effectively prevented apoptosis during porcine tetraploid embryo formation. Tetraploid aggregation-derived blastocysts have a high proportion of trophectoderm with increased expression of the CDX2 mRNA and high YAP1 intensity. High-quality blastocysts derived from a tetraploid embryo aggregation can serve as suitable source material for testing the differentiation potential of pluripotent stem cells for blastocyst complementation in pigs.

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Section: Discussionmentioning

confidence: 99%

Tetraploid embryo aggregation produces high-quality blastocysts with an increased trophectoderm in pigs

Lee,

Cai,

Kim

et al. 2023

Front. Cell Dev. Biol.

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“…Important events affecting cell reprogramming after SCNT include changes in chromatin accessibility [34], exchange of histone variants [35,36], silencing of genes expressed from the nuclear donor cell [30], DNA demethylation and remethylation [12], as well as changes in histone methylation [37] and histone acetylation patterns [38]. Among the attempts tested to enhance cell reprogramming, increasing histone acetylation has been shown to increase the development of SCNT in several species, including pigs [9,20]. Therefore, treatment with histone deacetylases inhibitors (HDACis), such as Scriptaid and Trichostatin A, has become an integral component of SCNT protocols.…”

Section: Discussionmentioning

confidence: 99%

“…Many approaches have been tested to overcome reprogramming roadblocks in SCNT embryos, such as chromatin and epigenetic regulators, which would create a more favorable environment for reprogramming [9]. Cellular reprogramming and totipotency acquisition are regulated by epigenetic modifications such as histone acetylation, as well as histone and DNA methylation [10,11].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, SCNT embryos were shown to have lower histone acetylation levels compared with embryos produced by in vitro fertilization (IVF) [15]. Thus, increasing histone acetylation by inhibiting histone deacetylases has been used to improve SCNT efficiency in several species, including pigs [9,[16][17][18][19]. Higher histone acetylation levels are related to a more open and permissive chromatin state, through reduction of the interaction between histones and DNA [20], which allows transcriptional activation and a better access of reprogramming factors to the chromatin.…”

Section: Introductionmentioning

confidence: 99%

“…Higher histone acetylation levels are related to a more open and permissive chromatin state, through reduction of the interaction between histones and DNA [20], which allows transcriptional activation and a better access of reprogramming factors to the chromatin. Scriptaid and Trichostatin A (TSA) are the more commonly used histone deacetylase inhibitors (HDACi) to improve cell reprogramming after SCNT in pig embryos [9]. Scriptaid seems to have lower cellular toxicity and promotes higher acetylation levels compared to other HDACi compounds [15,21].…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous Inhibition of Histone Deacetylases and RNA Synthesis Enables Totipotency Reprogramming in Pig SCNT Embryos

Macedo

Glanzner

Gutierrez

et al. 2022

IJMS

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Combining somatic cell nuclear transfer (SCNT) with genome editing technologies has emerged as a powerful platform for the creation of unique swine lineages for agricultural and biomedical applications. However, successful application of this research platform is still hampered by the low efficiency of these technologies, particularly in attaining complete cell reprogramming for the production of cloned pigs. Treating SCNT embryos with histone deacetylase inhibitors (HDACis), such as Scriptaid, has been routinely used to facilitate chromatin reprogramming after nuclear transfer. While increasing histone acetylation leads to a more relaxed chromatin configuration that facilitates the access of reprogramming factors and DNA repair machinery, it may also promote the expression of genes that are unnecessary or detrimental for normal embryo development. In this study, we evaluated the impact of inhibiting both histone deacetylases and RNA synthesis on pre- and post-implantation development of pig SCNT embryos. Our findings revealed that transcription can be inhibited for up to 40 h of development in porcine embryos, produced either by activation, fertilization or SCNT, without detrimentally affecting their capacity to form a blastocyst and their average number of cells at this developmental stage. Importantly, inhibiting RNA synthesis during HDACi treatment resulted in SCNT blastocysts with a greater number of cells and more abundant transcripts for genes related to embryo genome activation on days 2, 3 and 4 of development, compared to SCNT embryos that were treated with HDACi only. In addition, concomitant inhibition of histone deacetylases and RNA synthesis promoted the full reprograming of somatic cells, as evidenced by the normal fetal and full-term development of SCNT embryos. This combined treatment may improve the efficiency of the genome-editing + SCNT platform in swine, which should be further tested by transferring more SCNT embryos and evaluating the health and growth performance of the cloned pigs.

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Somatic Cell Nuclear Transfer in Pigs

Glanzner

Rissi

Bordignon

2023

Methods in Molecular Biology

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Enhancement of Chromatin and Epigenetic Reprogramming in Porcine SCNT Embryos—Progresses and Perspectives

Cited by 7 publications

References 150 publications

Tetraploid embryo aggregation produces high-quality blastocysts with an increased trophectoderm in pigs

Tetraploid embryo aggregation produces high-quality blastocysts with an increased trophectoderm in pigs

Simultaneous Inhibition of Histone Deacetylases and RNA Synthesis Enables Totipotency Reprogramming in Pig SCNT Embryos

Somatic Cell Nuclear Transfer in Pigs

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