FOXN1 基因敲除兔嵌合体的建立
作者:
作者单位:

1.五邑大学药学与食品工程学院,广东省医学大动物模型重点实验室,广东 江门 529020;2.广东省医学实验动物中心,南方医科大学附属广东省人民医院(广东省医学科学院),广东 佛山 528248

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R-33


Establishment of chimeric rabbits with FOXN1 gene knockout
Author:
Affiliation:

1. Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Pharmacy and Food Engineering,Wuyi University, Jiangmen 529020, China. 2. Guangdong Medical Laboratory Animal Center, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Foshan 528248

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    摘要:

    目的 建立 F0 代 FOXN1 基因敲除兔嵌合体,以探究普通饲养环境免疫缺陷兔活体保种的方法。 方法 首先,利用 CRISPR/ Cas9 技术,将构建好的 sgRNA 和 Cas9 蛋白注入兔二细胞期胚胎的一个细胞中,以获得FOXN1 基因编辑嵌合体胚胎。 然后,胚胎移植至代孕母兔。 最后,通过 PCR 技术以及 Sanger 测序方法鉴定 F0 代仔兔基因型,并观察其在普通饲养环境生长发育的情况。 结果 PCR 结合 Sanger 测序结果表明 FOXN1 基因敲除兔嵌合体构建成功。 经观察,嵌合体在普通环境下生长发育良好,无免疫缺陷表型。 结论 本研究初步建立了在普通环境下可正常生长发育的 FOXN1 基因敲除兔嵌合体,为后续进一步繁育 FOXN1 免疫缺陷兔奠定了研究基础。

    Abstract:

    Objective To establish F0 generation chimeric rabbits with FOXN1 gene knockout and explore method for the in vivo conservation of immunodeficient rabbits in a conventional housing environment. Methods Initially,CRISPR/ Cas9 technology was employed to inject constructed sgRNA and Cas9 protein into a single cell from rabbit two-cell stage embryos to obtain chimeric embryos with FOXN1 gene editing. The embryos were subsequently transferred into surrogate does. Finally, the F0 generation offsprings were genotyped using PCR and Sanger sequencing, and their growth and development in a conventional housing environment were observed. Results The PCR and Sanger sequencing result confirmed the successful establishment of chimeric rabbits with FOXN1 gene knockout. On observation, the chimeras exhibited normal growth and development in a conventional environment without any immunodeficient phenotypes. Conclusions This study established a preliminary chimeric rabbit model with FOXN1 gene knockout that grows and develops normally in standard laboratory environments. This lays the foundation for the further breeding of FOXN1 immunodeficient rabbits in the future.

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刘天平,黎桂玲,刘 科,陈傍柱,王 刚,顾为望.FOXN1 基因敲除兔嵌合体的建立[J].中国比较医学杂志,2024,34(4):35~40.

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  • 收稿日期:2023-11-15
  • 在线发布日期: 2024-06-20
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