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Generation of bicistronic reporter knockin mice for visualizing germ layers
http://hdl.handle.net/2241/00160022
f203c3b3-b8e6-444a-86a4-6ea334de8026
名前 / ファイル | ライセンス | Actions | |
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item type | Journal Article(1) | |||||||||||
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公開日 | 2020-05-12 | |||||||||||
タイトル | ||||||||||||
言語 | en | |||||||||||
タイトル | Generation of bicistronic reporter knockin mice for visualizing germ layers | |||||||||||
言語 | ||||||||||||
言語 | eng | |||||||||||
資源タイプ | ||||||||||||
タイプ | journal article | |||||||||||
著者 |
DINH, THI HUONG TRA
× DINH, THI HUONG TRA
WEKO
218873
× 村田, 知弥× 水野, 聖哉× 杉山, 文博× Suzuki, Hayate× Daitoku, Yoko× Tanimoto, Yoko× Kato, Kanako× Azami, Takuya× Ema, Masatsugu |
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抄録 | ||||||||||||
内容記述 | Knockout mouse models are commonly used in developmental biology to investigate the functions of specific genes, and the knowledge obtained in such models has yielded insights into the molecular mechanisms underlying developmental processes. Gastrulation is the most dynamic process in embryogenesis during which differentiation into three germ layers occurs. However, the functions of genes involved in gastrulation are not completely understood. One major reason for this is the technical difficulty of embryo analysis to understand germ layer location. We have generated three reporter mouse strains in which the germ layers are distinguished by different fluorescent reporters. Using CRISPR/Cas9 genome editing in mouse zygotes, the fluorescent reporter genes, EGFP, tdTomato, and TagBFP including 2A peptide sequences were knocked into the appropriate sites before the stop codon of the Sox17 (endoderm marker), Otx2 (ectoderm marker), and T (mesoderm marker) genes, respectively. Founder mice were successfully generated in the Sox17-2A-EGFP, Otx2-2A-tdTomato, and T-2A-TagBFP knockin reporter strains. Further, homozygous knockin mice of all strains appeared morphologically normal and were fertile. On stereomicroscopic analysis, fluorescent signals were detected in a germ layer-specific manner from heterozygous embryos at embryonic day (E) 6.5–8.5 in all strains, and were immunohistochemically demonstrated to match their respective germ layer-specific marker protein at E7.5. Taken together, these observations suggest that the Sox17-2A-EGFP, Otx2-2A-tdTomato, and T-2A-TagBFP knockin reporter mice may be useful for comprehensive analysis of gene function in germ layer formation. | |||||||||||
書誌情報 |
en : Experimental animals 巻 68, 号 4, p. 499-509, 発行日 2019-11 |
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ISSN | ||||||||||||
収録物識別子 | 1341-1357 | |||||||||||
書誌レコードID | ||||||||||||
収録物識別子 | AA11032321 | |||||||||||
PubMed番号 | ||||||||||||
関連識別子 | ||||||||||||
関連識別子 | 31189761 | |||||||||||
DOI | ||||||||||||
関連識別子 | ||||||||||||
関連識別子 | 10.1538/expanim.19-0031 | |||||||||||
権利 | ||||||||||||
権利情報 | ©2019 Japanese Association for Laboratory Animal Science | |||||||||||
権利 | ||||||||||||
権利情報 | This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License <http://creativecommons.org/licenses/by-nc-nd/4.0/>. | |||||||||||
著者版フラグ | ||||||||||||
値 | publisher | |||||||||||
出版者 | ||||||||||||
出版者 | Japanese Association for Laboratory Animal Science |