为探究tent5c/fam46c基因(Family with sequence similarity 46c,家族序列相似性46 c)在斑马鱼(Danio rerio)肌间刺发育中的作用,选取11种鱼类分析tent5c系统演化特征,采用原位杂交技术和qRT-PCR分析tent5c基因在斑马鱼发育中的时空表...为探究tent5c/fam46c基因(Family with sequence similarity 46c,家族序列相似性46 c)在斑马鱼(Danio rerio)肌间刺发育中的作用,选取11种鱼类分析tent5c系统演化特征,采用原位杂交技术和qRT-PCR分析tent5c基因在斑马鱼发育中的时空表达特征,并利用CRISPR/Cas9建立了斑马鱼tent5c基因敲除纯合突变系(tent5c^(-/-)),对其骨骼表型和肌肉转录组进行分析。结果显示,tent5c编码TENT5C氨基酸在不同鱼类中具有高度的保守性,但在无肌间刺鱼类里发生了相同的氨基酸替换;tent5c基因在48hpf(Hours post fertilization)斑马鱼的眼睛和肌隔表达,在60dpf(Days post fertilization)斑马鱼的脊椎和肌间刺特异高表达。与野生型斑马鱼相比,tent5c^(-/-)斑马鱼肌间刺表现为生长减缓,矿化程度降低以及数量减少。比较转录组结果表明,在tent5c基因敲除之后,斑马鱼肌肉组织有112个基因表达下调,118个基因表达上调。差异基因主要富集于肌肉收缩和骨骼发育等相关通路,由此推测tent5c可能是通过调控这些肌肉和骨骼相关通路来对肌间刺发育产生影响。研究初步表明了tent5c基因在斑马鱼肌间刺发育中的作用,为进一步研究鱼类肌间刺等骨骼发育的分子调控机制提供了科学依据。展开更多
The post-transcriptional regulation of mRNA is a crucial component of gene expression.The disruption of this process has detrimental effects on the normal development and gives rise to various diseases.Searching for n...The post-transcriptional regulation of mRNA is a crucial component of gene expression.The disruption of this process has detrimental effects on the normal development and gives rise to various diseases.Searching for novel post-transcriptional regulators and exploring their roles are essential for understanding development and disease.Through a multimodal analysis of red blood cell trait genome-wide association studies(GWAS)and transcriptomes of erythropoiesis,we identify FAM46C,a non-canonical RNA poly(A)polymerase,as a necessary factor for proper red blood cell development.FAM46C is highly expressed in the late stages of the erythroid lineage,and its developmental upregulation is controlled by an erythroidspecific enhancer.We demonstrate that FAM46C stabilizes mRNA and regulates erythroid differentiation in a polymerase activity-dependent manner.Furthermore,we identify transcripts of lysosome and mitochondria components as highly confident in vivo targets of FAM46C,which aligns with the need of maturing red blood cells for substantial clearance of organelles and maintenance of cellular redox homeostasis.In conclusion,our study unveils a unique role of FAM46C in positively regulating lysosome and mitochondria components,thereby promoting erythropoiesis.展开更多
文摘为探究tent5c/fam46c基因(Family with sequence similarity 46c,家族序列相似性46 c)在斑马鱼(Danio rerio)肌间刺发育中的作用,选取11种鱼类分析tent5c系统演化特征,采用原位杂交技术和qRT-PCR分析tent5c基因在斑马鱼发育中的时空表达特征,并利用CRISPR/Cas9建立了斑马鱼tent5c基因敲除纯合突变系(tent5c^(-/-)),对其骨骼表型和肌肉转录组进行分析。结果显示,tent5c编码TENT5C氨基酸在不同鱼类中具有高度的保守性,但在无肌间刺鱼类里发生了相同的氨基酸替换;tent5c基因在48hpf(Hours post fertilization)斑马鱼的眼睛和肌隔表达,在60dpf(Days post fertilization)斑马鱼的脊椎和肌间刺特异高表达。与野生型斑马鱼相比,tent5c^(-/-)斑马鱼肌间刺表现为生长减缓,矿化程度降低以及数量减少。比较转录组结果表明,在tent5c基因敲除之后,斑马鱼肌肉组织有112个基因表达下调,118个基因表达上调。差异基因主要富集于肌肉收缩和骨骼发育等相关通路,由此推测tent5c可能是通过调控这些肌肉和骨骼相关通路来对肌间刺发育产生影响。研究初步表明了tent5c基因在斑马鱼肌间刺发育中的作用,为进一步研究鱼类肌间刺等骨骼发育的分子调控机制提供了科学依据。
基金funded by the Starting Fund from Zhejiang University to N.L.and grants to X.L.from National Natural Science Foundation of China(82170120 and 81670108)CAMS Initiative for Innovative Medicine(2017-12M-B&R-04)+2 种基金Medical Epigenetics Research Cen-ter,CAMS(2018PT31015)the State Key Laboratory of Medical Molecular Biology(2060204)Haihe L aboratory of Cell Ecosystem Innovation Fund(22HHXBSS00008).
文摘The post-transcriptional regulation of mRNA is a crucial component of gene expression.The disruption of this process has detrimental effects on the normal development and gives rise to various diseases.Searching for novel post-transcriptional regulators and exploring their roles are essential for understanding development and disease.Through a multimodal analysis of red blood cell trait genome-wide association studies(GWAS)and transcriptomes of erythropoiesis,we identify FAM46C,a non-canonical RNA poly(A)polymerase,as a necessary factor for proper red blood cell development.FAM46C is highly expressed in the late stages of the erythroid lineage,and its developmental upregulation is controlled by an erythroidspecific enhancer.We demonstrate that FAM46C stabilizes mRNA and regulates erythroid differentiation in a polymerase activity-dependent manner.Furthermore,we identify transcripts of lysosome and mitochondria components as highly confident in vivo targets of FAM46C,which aligns with the need of maturing red blood cells for substantial clearance of organelles and maintenance of cellular redox homeostasis.In conclusion,our study unveils a unique role of FAM46C in positively regulating lysosome and mitochondria components,thereby promoting erythropoiesis.
