Quantitative spatial and temporal assessment of regulatory element activity in zebrafish.
Quantitative spatial and temporal assessment of regulatory element activity in zebrafish.
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DOI：
10.7554/elife.65601
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发表时间：
2021-11-19
期刊：
影响因子：
7.7
通讯作者：
Bickmore WA
中科院分区：
文献类型：
--
作者：
Bhatia S;Kleinjan DJ;Uttley K;Mann A;Dellepiane N;Bickmore WA
基因组非编码区的突变或遗传变异携带顺式调节元件（克雷斯），或增强子，已被广泛涉及人类疾病和疾病风险。然而，由于需要在适当的生物学背景下测定这些元素，我们测定这些DNA序列变化对增强子活性的影响的能力目前非常有限。在这里，我们描述了一种方法，同时定量评估的空间和时间活动的野生型和疾病相关的突变体人类CRE等位基因在斑马鱼胚胎发育中使用实时成像。我们产生了在斑马鱼基因组中预定义的中性对接位点中含有双CRE双报告基因盒的转基因系。每个CRE等位基因的活性通过特定荧光报告基因的表达来报告，从而允许同时可视化野生型等位基因在发育中何时何地是活性的以及该活性如何通过突变而改变。
Mutations or genetic variation in noncoding regions of the genome harbouring cis-regulatory elements (CREs), or enhancers, have been widely implicated in human disease and disease risk. However, our ability to assay the impact of these DNA sequence changes on enhancer activity is currently very limited because of the need to assay these elements in an appropriate biological context. Here, we describe a method for simultaneous quantitative assessment of the spatial and temporal activity of wild-type and disease-associated mutant human CRE alleles using live imaging in zebrafish embryonic development. We generated transgenic lines harbouring a dual-CRE dual-reporter cassette in a pre-defined neutral docking site in the zebrafish genome. The activity of each CRE allele is reported via expression of a specific fluorescent reporter, allowing simultaneous visualisation of where and when in development the wild-type allele is active and how this activity is altered by mutation.
在自失活逆转录病毒载体中组合掺入鸡 β 球蛋白 5'HS4 的增强子阻断成分和人 T 细胞受体 α/δ BEAD-1 绝缘子可降低其潜在的基因毒性。
影响因子：
5.2
作者：
Ramezani, Ali;Hawley, Teresa S.;Hawley, Robert G.
通讯作者：
Hawley, Robert G.