Photocaged morpholino oligomers for the light-regulation of gene function in zebrafish and Xenopus embryos.
Photocaged morpholino oligomers for the light-regulation of gene function in zebrafish and Xenopus embryos.
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DOI：
10.1021/ja1053863
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发表时间：
2010-11-10
影响因子：
15
通讯作者：
Yoder, Jeffrey A.
中科院分区：
文献类型：
--
作者：
Deiters, Alexander;Garner, R. Aaron;Lusic, Hrvoje;Govan, Jeane M.;Dush, Mike;Nascone-Yoder, Nanette M.;Yoder, Jeffrey A.
吗啉寡核苷酸，或吗啉寡核苷酸，已经成为在体外和体内环境中评估基因功能的强大的反义试剂。然而，这些试剂的构成活性限制了它们在需要时空控制的应用中的应用，例如胚胎中组织特异性基因的破坏。此外，目前间接调节体内吗啡诺活性的方法可能会产生非靶点效应。在这里，我们报道了一种新颖而有效的合成方法，将光笼中的单体构建块直接整合到吗啉低聚物中，并展示了这些笼养的吗啉在光激活控制细胞培养和活胚胎中的基因功能方面的应用。我们证明，仅在转基因细胞和活斑马鱼(Danio Rerio)和非洲爪蛙胚胎暴露于紫外线后，以增强绿色荧光蛋白(EGFP)为靶标的笼式吗啉才会干扰EGFP的产生。最后，我们发现，以Chordin为靶标的笼式吗啉以UV依赖的方式诱导Chordin表型。Chordin是一种斑马鱼基因，当被传统的吗啉干扰时，会产生不同的表型。我们的结果表明，直接光笼吗啉是一种容易合成的高效工具，可以在多个环境中进行光激活的基因表达时空调控。
Morpholino oligonucleotides, or morpholinos, have emerged as powerful antisense reagents for evaluating gene function in both in vitro and in vivo contexts. However, the constitutive activity of these reagents limits their utility for applications that require spatiotemporal control, such as tissue specific gene disruptions in embryos. In addition, current indirect methods for spatiotemporal regulation of morpholino activity in vivo may have off-target effects. Here we report a novel and efficient synthetic route for directly incorporating photocaged monomeric building blocks into morpholino oligomers, and demonstrate the utility of these caged morpholinos in the light-activated control of gene function in both cell culture and living embryos. We demonstrate that a caged morpholino targeting enhanced green fluorescent protein (EGFP) disrupts EGFP production only after exposure to UV light in both transfected cells and living zebrafish (Danio rerio) and Xenopus frog embryos. Finally, we show that a caged morpholino targeting chordin, a zebrafish gene that yields a distinct phenotype when functionally disrupted by conventional morpholinos, elicits a chordin phenotype in a UV-dependent manner. Our results suggest that directly photocaged morpholinos are readily synthesized and highly efficacious tools for light-activated spatio-temporal control of gene expression in multiple contexts.
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影响因子：
48
作者：
Cambridge, Sidney B.;Geissler, Daniel;Bonhoeffer, Tobias
通讯作者：
Bonhoeffer, Tobias
影响因子：
--
作者：
Blidner, Richard A.;Svoboda, Kurt R.;Monroe, W. Todd
通讯作者：
Monroe, W. Todd
影响因子：
5.2
作者：
Lusic, Hrvoje;Young, Douglas D.;Deiters, Alexander
通讯作者：
Deiters, Alexander
DOI：
10.1089/oli.1.1997.7.151
发表时间：
1997-06-01
期刊：
ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT
影响因子：
--
作者：
Stein, D;Foster, E;Summerton, J
通讯作者：
Summerton, J
影响因子：
15
作者：
Ouyang X;Shestopalov IA;Sinha S;Zheng G;Pitt CL;Li WH;Olson AJ;Chen JK
通讯作者：
Chen JK