胚皮细胞和CRISPR/CAS9技术在鸭转基因中的应用

Q4 Biochemistry, Genetics and Molecular Biology

Biopolymers and Cell Pub Date : 2021-08-31 DOI:10.7124/bc.000a5b

O. Konoval, P. Korol, S. Kostenko, P. Tabaka, L. Lizhi, A. Chepiha, M. Doroshenko, O. Sydorenko, P. Dzhus, N. Svyrydenko, T. Lytvynenko, B. Xingchen, H. Xuetao, L. Li, M. Drahulian, E. Kostyuk, P. Filipova

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引用次数: 0

摘要

本研究的目的是通过DNA载体和Lipofectamine 2000®(Invitrogen公司，美国)转染供体胚皮细胞，建立一种鸭子转基因嵌合体的方法。采用同源定向修复(homology directed repair, HDR) CRISPR/Cas9系统编辑鸭基因组靶位点。材料和研究方法。用质粒DNA和Lipofectamine 2000转染供体胚皮细胞，建立了转基因鸭嵌合体。为了编辑鸭基因组的目标区域，我们使用了带有HDR的CRISPR / cas9系统。采用EGFP报告基因作为转基因。阳性(30.3%)，其中男性10例(16.9%)，女性49例(83.1%)。我们所使用的技术可以成功地应用于进一步的研究和转基因鸭的创造。

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Efficacy of blastodermal cells and CRISPR/CAS9 method in the creation of transgenic duck (Anas Platyrhynchos)

The aim of the work was to develop a methodology for the creation of transgenic chimeras of ducks by using donor blastodermal cells after transfection with DNA vector and Lipofectamine 2000® (Invitrogen, USA). The CRISPR/Cas9 system with homology directed repair (HDR) was used to edit the target site of the duck genome. Materials and research methods. Transgenic duck chimeras were created using donor blastodermal cells after transfection with plasmid DNA and Lipofectamine 2000. To edit the target region of the duck genome, we used the CRISPR / cas9 system with HDR. The EGFP reporter gene was used as the transgene. positive (30.3 %), including 10 males (16.9 %) and 49 females (83.1 %). The technique used by us can be successfully applied in further researches and for creation of a transgenic duck.

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来源期刊

Biopolymers and Cell Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)

CiteScore

1.10

自引率

0.00%

发文量

期刊介绍： “Biopolymer and cell” is published since 1985 at the Institute of Molecular Biology and Genetics NAS of Ukraine under the supervision of the National Academy of Sciences of Ukraine. Our journal covers a wide scope of problems related to molecular biology and genetics including structural and functional genomics, transcriptomics, proteomics, bioinformatics, biomedicine, molecular enzymology, molecular virology and immunology, theoretical bases of biotechnology, physics and physical chemistry of proteins and nucleic acids and bioorganic chemistry.