Afp-cre-lacZ转基因小鼠的构建

doi:10.3969/j.issn.1674-5817.2017.02.002

实验动物与比较医学 ›› 2017, Vol. 37 ›› Issue (2): 89-93.DOI: 10.3969/j.issn.1674-5817.2017.02.002

所属专题：实验动物资源开发与利用

Afp-cre-lacZ转基因小鼠的构建

顾晓雯¹, 孙瑞林², 费俭¹

1.同济大学生命科学与技术学院, 上海 200092;
2.上海南方模式生物科技股份有限公司, 上海 201203

收稿日期:2017-02-05 出版日期:2017-04-25 发布日期:2017-04-25
作者简介:顾晓雯(1988-),女,博士,从事生物化学与分子生物学研究。E-mail:17guxiaowen@tongji.edu.cn
基金资助:
国家重点基础研究发展计划(2010CB945501)

Construction of Afp-cre-lacZ Transgenic Mouse Model

GU Xiao-wen¹, SUN Rei-lin², FEI Jian¹

1. School of Life Science and Technology, Tongji University, Shanghai 200092, China;
2. Shanghai Biomodel Organism Science & Technology Development Co.,Ltd., Shanghai 201203, China

Received:2017-02-05 Published:2017-04-25 Online:2017-04-25

摘要/Abstract

摘要： 目的研究甲胎蛋白(Afp)阳性细胞的增殖分化在组织生长修复,肿瘤发生过程中的作用,建立Afp-CreERT转基因小鼠细胞示踪系统。方法采用DNA雄原核显微注射方法获得Afp-CreERT转基因小鼠,筛选出合适的品系后,使之与Rosa26-lacZ工具小鼠杂交,获得Cre/lacZ双阳性的Afp-cre-lacZ转基因小鼠。结果显微注射后,共出生小鼠56只,经PCR鉴定Cre阳性小鼠共4只,阳性小鼠传代后各为一系。筛选内源性Afp表达与Cre表达相对符合的品系作为Afp-CreERT转基因小鼠品系建系。Afp-CreERT转基因小鼠与Rosa26-lacZ工具小鼠杂交,经PCR鉴定后获得Cre/lacZ双阳性的Afp-cre-lacZ转基因小鼠。经实时PCR,X-gal染色,免疫荧光染色鉴定后得到Afp-cre-lacZ转基因小鼠细胞示踪系统,同时证实该系统能够正确示踪Afp表达阳性的细胞,同时也能够应用于肝损伤小鼠模型中。结论成功构建了Afp-cre-lacZ转基因小鼠细胞示踪系统,为研究这类细胞的谱系发生提供了工具。

关键词: 甲胎蛋白(Afp), 转基因小鼠模型, 细胞示踪

Abstract: Objective To study the function of alpha-fetoprotein (Afp) positive cells during tissues growth and repair or in tumorigenesis, Afp-cre-lacZ linage tracing mouse model was established. Methods Afp-CreERT mice were established by microinjection and identified by real time PCR. Afp-CreERT mice were crossed with Rosa26-lacZ mice. Cre/lacZ both positive mice were kept as Afp-cre-lacZ mice. Results Fifty-six transgenic mice were born after microinjection. Four of them genotyped Cre positive were kept as founder. Each founder was established a strain. Cre expression of each strain was identified via real time PCR. Thus Afp-CreERT transgenic mouse was established. The Cre/lacZ both positive offspring of Afp-CreERT mice were crossed with Rosa26-lacZ mice were kept as Afp-cre-lacZ mice. Identified by real time PCR, X-gal staining and immunofluorescence, Afp-cre-lacZ mice were observed that they could be used in lineage tracing and further studies. Conclusions The Afp-cre-lacZ mice established may be used as a tool for lineage tracing studies.

Key words: Alpha-fetoprotein (Afp), Transgenic mouse model, Lineage tracing

中图分类号:

Q95-33

顾晓雯,孙瑞林,费俭. Afp-cre-lacZ转基因小鼠的构建[J]. 实验动物与比较医学, 2017, 37(2): 89-93. DOI: 10.3969/j.issn.1674-5817.2017.02.002.

GU Xiao-wen,SUN Rei-lin,FEI Jian. Construction of Afp-cre-lacZ Transgenic Mouse Model[J]. Laboratory Animal and Comparative Medicine, 2017, 37(2): 89-93. DOI: 10.3969/j.issn.1674-5817.2017.02.002.

参考文献

[1] Sun W, Liu Y, Shou D, et al.AFP (alpha fetoprotein): who are you in gastrology?[J]. Cancer Lett, 2015, 357(1):43-46.
[2] Terentiev AA, Moldogazieva NT.Alpha-fetoprotein: a renaissance[J]. Tumour Biol, 2013, 34(4):2075-2091.
[3] Kajiyama Y, Tian J, Locker J.Regulation of alpha-fetoprotein expression by Nkx2.8[J]. Mol Cell Biol, 2002, 22(17):6122-6130.
[4] Peterson ML, Ma C, Spear BT.Zhx2 and Zbtb20: novel regulators of postnatal alpha-fetoprotein repression and their potential role in gene reactivation during liver cancer[J]. Semin Cancer Biol, 2011, 21(1):21-27.
[5] Nakao K, Ichikawa T.Recent topics on alpha-fetoprotein[J]. Hepatol Res, 2013, 43(8):820-825.
[6] Forner A, Llovet JM, Bruix J.Hepatocellular carcinoma[J]. Lancet, 2012, 379(9822):1245-1255.
[7] Song PP, Xia JF, Inagaki Y, et al.Controversies regarding and perspectives on clinical utility of biomarkers in hepatocellular carcinoma[J]. World J Gastroenterol, 2016, 22(1):262-274.
[8] Tang H, Tang XY, Liu M, et al.Targeting alpha-fetoprotein represses the proliferation of hepatoma cells via regulation of the cell cycle[J]. Clin Chim Acta, 2008, 394(1):81-88.
[9] Parpart S, Roessler S, Dong F, et al.Modulation of miR-29 expression by alpha-fetoprotein is linked to the hepatocellular carcinoma epigenome[J]. Hepatology, 2014, 60(3):872-883.
[10] Chen L, Zhang W, Liang HF, et al.Activin A induces growth arrest through a SMAD-dependent pathway in hepatic progenitor cells[J]. Cell Commun Signal, 2014, 12:18.
[11] Zhao BB, Li HM, Gao X, et al.The herbal compound "diwu yanggan" modulates liver regeneration by affecting the hepatic stem cell microenvironment in 2-acetylaminofluorene/partial hepatectomy rats[J]. Evid Based Complement Alternat Med, 2015, 2015:468303.
[12] Sauzay C, Petit A, Bourgeois AM, et al.Alpha-foetoprotein (AFP): A multi-purpose marker in hepatocellular carcinoma[J]. Clin Chim Acta, 2016, 463:39-44.
[13] Wang B, Zhao L, Fish M, et al.Self-renewing diploid Axin2(+) cells fuel homeostatic renewal of the liver[J]. Nature, 2015, 524(7564):180-185.
[14] Tarlow BD, Finegold MJ, Grompe M.Clonal tracing of Sox9+ liver progenitors in mouse oval cell injury[J]. Hepatology, 2014, 60(1):278-289.
[15] Malato Y, Naqvi S, Schurmann N, et al.Fate tracing of mature hepatocytes in mouse liver homeostasis and regeneration[J]. J Clin Invest, 2011, 121(12):4850-4860.
[16] Shin S, Upadhyay N, Greenbaum LE, et al.Ablation of Foxl1-Cre-labeled hepatic progenitor cells and their descendants impairs recovery of mice from liver injury[J]. Gastroenterology, 2015, 148(1):192-202.

Afp-cre-lacZ转基因小鼠的构建

Construction of Afp-cre-lacZ Transgenic Mouse Model

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