Evaluating histone modification analysis of individual preimplantation embryos
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- 曾, 毅仁
- Laboratory of Reproductive Biology, Graduate School of Agriculture
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- 須佐見, 和生
- Laboratory of Reproductive Biology, Graduate School of Agriculture
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- 星野, 洋一郎
- Laboratory of Reproductive Biology, Graduate School of Agriculture
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- 本多, 慎之介
- Laboratory of Reproductive Biology, Graduate School of Agriculture
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- 南, 直治郎
- Laboratory of Reproductive Biology, Graduate School of Agriculture
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- 池田, 俊太郎
- Laboratory of Reproductive Biology, Graduate School of Agriculture
説明
[Background] We previously reported a modification of the CUT&Tag method (NTU-CAT) that allows genome-wide histone modification analysis in individual preimplantation embryos. In the present study, NTU-CAT was further simplified by taking advantage of the Well-of-the-Well (WOW) system, which enables the processing of multiple embryos in a shorter time with less reagent and cell loss during the procedure (WOW-CUT&Tag, WOW-CAT). [Results] WOW-CAT allowed histone modification profiling from not only a single blastocyst but also from a portion of it. WOW-CAT generated similar H3K4me3 profiles as NTU-CAT, but they were closer to the profiles produced by chromatin immunoprecipitation-sequencing, such as a valley-like trend and relatively lower false positive rates, indicating that WOW-CAT may attenuate the bias of Tn5 transposase to cut open chromatin regions. Simultaneous WOW-CAT of two halves of single blastocysts was conducted to analyze two different histone modifications (H3K4me3 and H3K27ac) within the same embryo. Furthermore, trophectoderm cells were biopsied and subjected to WOW-CAT in anticipation of preimplantation diagnosis of histone modifications. WOW-CAT allowed the monitoring of epigenetic modifications in the main body of the embryo. For example, analysis of H3K4me3 modifications of XIST and DDX3Y in trophectoderm biopsies could be used to sex embryos in combination with quantitative PCR, but without the need for deep sequencing. [Conclusions] These results suggest the applicability of WOW-CAT for flexible epigenetic analysis of individual embryos in preimplantation epigenetic diagnosis.
収録刊行物
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- BMC Genomics
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BMC Genomics 25 2024-01-18
Springer Nature
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詳細情報 詳細情報について
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- CRID
- 1050580450345294976
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- ISSN
- 14712164
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- HANDLE
- 2433/286756
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- 本文言語コード
- en
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- 資料種別
- journal article
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- データソース種別
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- IRDB