Dong Xiaohua, Li Jingyun, He Luqingqing, Gu Chun, Jia Wenshuang, Yue Yunyun, Li Jun, Zhang Qinxin, Chu Lele, Zhao Qingshun
Ministry of Education Key Laboratory of Model Animal for Disease Study, Model Animal Research Center, Nanjing University, Nanjing 210061, China; Institute of Genome Editing, Nanjing YSY Biotech Company, Limited, Nanjing 211812, China.
Maternal and Child Health Medical Institute, Department of Plastic and Cosmetic Surgery, Obstetrics and Gynecology Hospital Affiliated to Nanjing Medical University, Nanjing 210004, China.
J Biol Chem. 2017 Aug 4;292(31):13045-13055. doi: 10.1074/jbc.M117.777094. Epub 2017 Jun 16.
Transcription factors play crucial roles in patterning posterior neuroectoderm. Previously, zinc finger transcription factor was reported to be expressed in the posterior neuroectoderm of zebrafish embryos. However, its roles remain unknown. Here, we report that there are 13 copies of in the zebrafish genome, and all the paralogues share highly identical protein sequences and cDNA sequences. When are knocked down using a morpholino to inhibit their translation or dCas9-Eve to inhibit their transcription, the zebrafish gastrula displays reduced expression of , the marker gene for the posterior neuroectoderm. Further analyses reveal that diminishing produces the decreased expressions of , whereas overexpression of effectively rescues the reduced expression of in the posterior neuroectoderm. Additionally, knocking down causes the reduced expression of , a direct regulator of , in the posterior neuroectoderm, and overexpression of rescues the expression of in the knockdown embryos. In contrast, knocking down either or does not affect the expressions of Taken together, our results demonstrate that zebrafish control the expression of in the posterior neuroectoderm by acting upstream of and .
转录因子在后部神经外胚层的模式形成中发挥着关键作用。此前，据报道锌指转录因子在斑马鱼胚胎的后部神经外胚层中表达。然而，其作用仍不清楚。在此，我们报道在斑马鱼基因组中有13个该基因的拷贝，并且所有的旁系同源物都具有高度相同的蛋白质序列和cDNA序列。当使用吗啉代寡核苷酸抑制其翻译或dCas9-Eve抑制其转录来敲低该基因时，斑马鱼原肠胚中后部神经外胚层的标记基因的表达降低。进一步分析表明，减少该基因的表达会导致相关基因的表达下降，而该基因的过表达有效地挽救了后部神经外胚层中相关基因表达的降低。此外，敲低该基因会导致后部神经外胚层中该基因的直接调节因子的表达降低，而该基因的过表达挽救了敲低胚胎中该调节因子的表达。相反，敲低该基因或该基因均不影响相关基因的表达。综上所述，我们的结果表明斑马鱼的该基因通过在该基因和该基因的上游起作用来控制后部神经外胚层中该基因的表达。
