SD

针对线粒体DNA G7755位点设计构建DdCBE载体，利用显微注射将DdCBE工具导入大鼠受精卵，然后移植到假孕大鼠中（见下图）。

经抓力和转棒测试，线粒体DNA TRNK G7755A点突变大鼠的抓力和运动能力相比野生对照大鼠无显著变化。 多普勒超声检测结果显示，G7755A点突变大鼠的左心室舒张末和收缩末期内径无显著变化；舒张末和收缩末时左室前壁厚度无显著改变；左心室射血分数以及左室缩短百分比无显著变化。

Fig. 1 Mitochondrial disorder modeling by DdCBE in rat. a Representative sequence chromatograms of G7755A (Left) and G14098A (Right)founder rats. Target sites are indicated by red arrows. b Frequencies of G-to-A conversion at G7755 (Blue dots) and G14098 (Red dots) in founder rats. c Frequencies of mtDNA G-to-A conversion of offspring from G14098A #7 founder. The red dot indicates founder, black dots indicate F1 offspring. d Measuring distance (Left) and speed (Right) by open field test. e Evaluation of motor coordination by Rotarod test. f Measuring grip strength by GripStrength Test. g–m Echocardiography analysis of wild-type rats and edited F1 males. The tests include a snapshot of M-mode of echocardiography(g), left ventricular (LV) diameter at end-diastole and end-systole (LVIDD, LVIDS) (h, i), LV anterior wall thickness at end-diastole and end-systole(LVAWD, LVAWS) (j, k), LV ejection fraction (LVEF) (l), and LV percent fractional shortening (LVFS) (m). Data were presented as a scatter dot plot withmeans (n = 4 for wild-type control, n = 5 for G14098A F1 rats). Significance was calculated with unpaired two-tailed Student’s t-test(*P < 0.05, **P < 0.01, ***P < 0.001)

利用DdCBE工具构建的G7755A线粒体点突变大鼠为相关疾病研究提供大鼠模型。