Evaluation of developmental phenotypes produced by morpholino antisense targeting of a sea urchin Runx gene.
Evaluation of developmental phenotypes produced by morpholino antisense targeting of a sea urchin Runx gene.
复制标题
评估吗啉代反义靶向海胆 Runx 基因产生的发育表型。
DOI：
10.1186/1741-7007-2-6
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发表时间：
2004-05-07
期刊：
影响因子：
5.4
通讯作者：
Robertson, Anthony J.
中科院分区：
文献类型：
--
作者：
Coffman, James A.;Dickey-Sims, Carrie;Haug, Jeffrey S.;McCarthy, John J.;Robertson, Anthony J.
Runx 转录因子是后生动物发育的重要调节因子。海胆 Runx 基因 SpRunt 先前被鉴定为 CyIIIa 肌动蛋白基因的反式激活因子，CyIIIa 肌动蛋白基因是幼虫非外胚层的分化标记。在这里，我们扩展了 SpRunt 的功能分析，使用吗啉代反义寡核苷酸（吗啉代）来干扰胚胎中 SpRunt 的表达。评估了四种不同 SpRunt 特异性吗啉代的发育效应。最有效地敲低 SpRunt 的两种吗啉代在裂解后期产生相同的有丝分裂灾难表型，这是巧合错误靶向组蛋白 mRNA 的产物，为两种不同吗啉代作为特异性对照的不足提供了一个警示性的例子。另外两种吗啡啉会产生原肠胚阶段的增殖和分化缺陷，这些缺陷可以通过外源 SpRunt mRNA 来挽救。通过定量聚合酶链反应分析了后期胚胎中涉及细胞增殖和分化的 22 个基因的表达。研究发现，SpRunt 的敲低会扰乱所有主要组织区域中分化标记物的表达以及细胞周期控制基因（包括细胞周期蛋白 B 和细胞周期蛋白 D）的表达。SpRunt 对于胚胎发育至关重要，并且是协调细胞增殖和分化的全局必需的。
Runx transcription factors are important regulators of metazoan development. The sea urchin Runx gene SpRunt was previously identified as a trans-activator of the CyIIIa actin gene, a differentiation marker of larval aboral ectoderm. Here we extend the functional analysis of SpRunt, using morpholino antisense oligonucleotides (morpholinos) to interfere with SpRunt expression in the embryo. The developmental effects of four different SpRunt-specific morpholinos were evaluated. The two morpholinos most effective at knocking down SpRunt produce an identical mitotic catastrophe phenotype at late cleavage stage that is an artifact of coincidental mis-targeting to histone mRNA, providing a cautionary example of the insufficiency of two different morpholinos as a control for specificity. The other two morpholinos produce gastrula stage proliferation and differentiation defects that are rescued by exogenous SpRunt mRNA. The expression of 22 genes involved in cell proliferation and differentiation was analyzed in the latter embryos by quantitative polymerase chain reaction. Knockdown of SpRunt was found to perturb the expression of differentiation markers in all of the major tissue territories as well as the expression of cell cycle control genes, including cyclin B and cyclin D. SpRunt is essential for embryonic development, and is required globally to coordinate cell proliferation and differentiation.
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影响因子：
1.5
作者：
Draper, BW;Morcos, PA;Kimmel, CB
通讯作者：
Kimmel, CB
影响因子：
10.5
作者：
Sullivan, E;Santiago, C;Duronio, RJ
通讯作者：
Duronio, RJ
影响因子：
2.7
作者：
Cavalieri, V;Spinelli, G;Di Bernardo, M
通讯作者：
Di Bernardo, M
影响因子：
2.7
作者：
Davidson, EH;Rast, JP;Bolouri, H
通讯作者：
Bolouri, H
影响因子：
5.3
作者：
Moore, JC;Sumerel, JL;Marzluff, WF
通讯作者：
Marzluff, WF