Oncotarget

Research Papers:

Generation of TALEN-mediated FH knockout rat model

Dandan Yu, Yali Zhong, Xiaoran Li, Yaqing Li, Xiaoli Li, Jing Cao, Zhirui Fan, Huijie Fan, Long Yuan, Benling Xu, Yuan Yuan, Hongquan Zhang, Zhenyu Ji, Jian-Guo Wen, Mingzhi Zhang, Jahn M. Nesland and Zhenhe Suo _

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Oncotarget. 2016; 7:61656-61669. https://doi.org/10.18632/oncotarget.11429

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Abstract

Dandan Yu1,9,10, Yali Zhong1, Xiaoran Li9,10, Yaqing Li1, Xiaoli Li1, Jing Cao1,2, Zhirui Fan1, Huijie Fan1, Long Yuan3, Benling Xu4, Yuan Yuan5, Hongquan Zhang6, Zhenyu Ji7, Jian-Guo Wen8, Mingzhi Zhang1, Jahn M. Nesland9,10, Zhenhe Suo1,9,10

1Department of Oncology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China

2Departments of Pathology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China

3Department of Surgery, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan, China

4Department of Cancer Biotherapy, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan, China

5Department of Pathology, Capital Medical University, Beijing, Fengtai, China

6Department of Anatomy, Histology and Embryology, Peking University Health Science Center, Beijing, Fengtai, China

7Department of Oncology, Henan Academy of Medical and Pharmaceutical Sciences, Zhengzhou University, Zhengzhou, Henan, China

8Institute of Clinical Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou University, Zhengzhou, Henan, China

9Department of Pathology, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway

10Department of Pathology, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway

Correspondence to:

Zhenhe Suo, email: [email protected]

Keywords: TALEN, fumarate hydratase, gene technology, obesity, heterozygote

Received: February 23, 2016     Accepted: August 09, 2016     Published: August 19, 2016

ABSTRACT

Transcription activator-like effector nucleases (TALENs) are valuable tools for precise genome engineering of laboratory animals. Here we utilized this technique for efficient site-specific gene modification to create a fumarate hydratase (FH) gene knockout rat model, in which there was an 11 base-pair deletion in the first exon of the FH gene in 111 rats. 18 live-born targeted mutation offsprings were produced from 80 injected zygotes with 22.5% efficiency, indicating high TALEN knockout success in rat zygots. Only heterozygous deletion was observed in the offsprings. Sixteen pairs of heterozygous FH knockout (FH+/–) rats were arranged for mating experiments for six months without any homozygous KO rat identified. Sequencing from the pregnant rats embryo samples showed no homozygous FH KO, indicating that homozygous FH KO is embryonically lethal. Comparatively, the litter size was decreased in both male and female FH+/– KO rats. There was no behaviour difference between the FH+/– KO and the control rats except that the FH+/– KO male rats showed significantly higher body weight in the 16-week observation period. Clinical haematology and biochemical examinations showed hematopoietic and kidney dysfunction in the FH+/– KO rats. Small foci of anaplastic lesions of tubular epithelial cells around glomeruli were identified in the FH+/– kidney, and these anaplastic cells were comparatively positive for Ki67, p53 and Sox9, and such findings are most probably related to the kidney dysfunction reflected by the biochemical examinations of the rats. In conclusion, we have successfully established an FH+/– KO rat model, which will be useful for further functional FH studies.


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