关键词: TALEN基因敲除 OGDH SD大鼠 瓦博格效应

Abstract: Objective: To establish OGDH gene knockout SD rats by transcription activator-like effector nucleases (TALEN) technology. Methods: Three pairs of TALEN plasmid were designed and constructed, then the transfection activity was examined in 293T cells, and the TALEN pair with the most effective mutation result was selected before the plasmid pair was applied to the fertilized rat eggs using microinjection. DNA sequencing with tissue samples from two-weeks newborn SD rats were performed. The SD rats were followed by measuring the body weight, observing food taking, running activity and fur color etc. Theprotein expression of OGDH was detected by Western Blot. Results: The TALEN plasmid pair OGDH-T2 was found the most effective and this plasmid pair was used in this study.In total, 80 fertilized eggs were injected with the OGDH-T2 plasmid pair and 18 rats were born, with an efficiency rate of 22.5%. However, homogeneous knockout rats were notobtained after followed a period of three generations. However, heterogeneous KO mutation OGDH-KO8 could be consistently maintained. Therefore, we followed this type of heterogeneous KO rats in this study. We discovered that OGDH-KO8 influenced the bodyweight. Compared with the control rats, the rats with the OGDH-KO8 heterogeneous mutation showed significantly higher body weight within the 10 weeks′observation(P<0.001), Otherwise, there was no difference observed in this typical heterogeneous KO rats. In consistence with the bodyweight increase, the OGDH protein expression was decreased in theserats, the rates of decreased up to 50.6%(P<0.01), the expression of liver OGDH protein is lower 50.6% than the WT type.Conclusion: Heterogeneous OGDH gene knockout SD rats are successfully established.

Key words: TALEN, gene knockout, OGDH, SD rats, Warburg effect