利用CRISPR/Cas9基因编辑技术在斑马鱼中构建了可稳定遗传的生长抑素(somatostatin,SST)基因(sst1)突变体鱼系。通过比较6 dpf(days post fertilization,dpf)的sst1^(-/-)突变体和野生型仔鱼转录组,发现sst1^(-/-)突变体相关生化过程和...利用CRISPR/Cas9基因编辑技术在斑马鱼中构建了可稳定遗传的生长抑素(somatostatin,SST)基因(sst1)突变体鱼系。通过比较6 dpf(days post fertilization,dpf)的sst1^(-/-)突变体和野生型仔鱼转录组,发现sst1^(-/-)突变体相关生化过程和信号通路发生了显著变化。相比野生型,sst1^(-/-)突变体仔鱼中有354个基因显著上调,504个基因显著下调。GO(gene ontology)富集分析表明,大部分差异基因与氨基酸和蛋白质生物合成相关。KEGG(kyoto encyclopedia of genes and genomes)通路富集显示代谢通路、氨基酸合成、氨基酰基-tRNA合成、核糖体合成和rRNA加工相关通路表达增强。结果表明,sst1^(-/-)突变体蛋白质生物合成和代谢过程更加活跃。RT-qPCR验证结果显示,随机选取的10个基因mRNA表达变化趋势与转录组测序结果一致,说明转录组测序结果真实可靠。6月龄突变体体质量、体长和野生型相比并无显著差异。本研究结果为进一步挖掘生长抑素基因家族的潜在功能奠定基础。展开更多
The G protein coupled receptor（GPCR）, one of the members in the superfamily, which consists of thousands of integral membrane proteins, exerts a wide variety of physiological functions and responses to a large porti...The G protein coupled receptor（GPCR）, one of the members in the superfamily, which consists of thousands of integral membrane proteins, exerts a wide variety of physiological functions and responses to a large portion of the drug targets. The 3D structure of somatostatin receptor I（SSTR1） was modeled and refined by means of homology modeling and molecular dynamics simulation. This model was assessed by Verify-3D and Vadar, which confirmed the reliability of the refined model. The interaction between the inhibitor cysteamine, somatostatin（SST） and SSTR1 was investigated by a molecular docking program, Affinity. The binding module not only showed the crucial residues involved in the interaction, but also provided important information about the interaction between SSTR1 on the one hand and ligands on the other, which might be the significant evidence for the structure-based design.展开更多
文摘利用CRISPR/Cas9基因编辑技术在斑马鱼中构建了可稳定遗传的生长抑素(somatostatin,SST)基因(sst1)突变体鱼系。通过比较6 dpf(days post fertilization,dpf)的sst1^(-/-)突变体和野生型仔鱼转录组,发现sst1^(-/-)突变体相关生化过程和信号通路发生了显著变化。相比野生型,sst1^(-/-)突变体仔鱼中有354个基因显著上调,504个基因显著下调。GO(gene ontology)富集分析表明,大部分差异基因与氨基酸和蛋白质生物合成相关。KEGG(kyoto encyclopedia of genes and genomes)通路富集显示代谢通路、氨基酸合成、氨基酰基-tRNA合成、核糖体合成和rRNA加工相关通路表达增强。结果表明,sst1^(-/-)突变体蛋白质生物合成和代谢过程更加活跃。RT-qPCR验证结果显示,随机选取的10个基因mRNA表达变化趋势与转录组测序结果一致,说明转录组测序结果真实可靠。6月龄突变体体质量、体长和野生型相比并无显著差异。本研究结果为进一步挖掘生长抑素基因家族的潜在功能奠定基础。
基金the National Basic Research Program of China，the National Natural Science Foundation of China
文摘The G protein coupled receptor（GPCR）, one of the members in the superfamily, which consists of thousands of integral membrane proteins, exerts a wide variety of physiological functions and responses to a large portion of the drug targets. The 3D structure of somatostatin receptor I（SSTR1） was modeled and refined by means of homology modeling and molecular dynamics simulation. This model was assessed by Verify-3D and Vadar, which confirmed the reliability of the refined model. The interaction between the inhibitor cysteamine, somatostatin（SST） and SSTR1 was investigated by a molecular docking program, Affinity. The binding module not only showed the crucial residues involved in the interaction, but also provided important information about the interaction between SSTR1 on the one hand and ligands on the other, which might be the significant evidence for the structure-based design.
