High-efficiency gene transfer into adult fish: A new tool to study fin regeneration
High-efficiency gene transfer into adult fish: A new tool to study fin regeneration
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DOI：
10.1002/gene.10025
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发表时间：
2002-01-01
期刊：
影响因子：
1.5
通讯作者：
Paulin, D
中科院分区：
文献类型：
--
作者：
Tawk, M;Tuil, D;Paulin, D
斑马鱼代表了研究脊椎动物基因功能的极好模型（例如，发达的遗传学，大量的突变体，以及正在进行的基因组测序），因为我们有工具来操纵基因表达。最近在卵中注射的吗啉代的使用提供了在早期发育中“敲低”11个基因表达的良好方法（Nasevicius和Ekker，2000），并且在卵中注射的“笼状”RNA允许在特定组的细胞中过表达基因（Ando等人，2001）。然而，在青少年或成年人中特异性修饰基因表达的方法仍然缺失。这种方法将是了解分化组织中基因功能的一个非常强大的工具。我们在这里描述了一种基于电穿孔的方法，该方法允许在成人组织中进行基因转移。使用GFP（绿色荧光蛋白）依赖性测定评估其效率。然后，我们使用这种方法来破坏再生过程中的FGF信号通路。(C)2002 Wiley-Liss，Inc.
Zebrafish represents an excellent model to study the function of vertebrate genes (e.g., well-developed genetics, large number of mutants, and genomic sequencing in progress), inasmuch as we have tools to manipulate gene expression. Recent use of injected morpholinos in eggs provides a good method to '' knockdown 11 gene expression in early development, (Nasevicius and Ekker, 2000), and the ''caged'' RNA injected in eggs allows to overexpress a gene in a specific set of cells (Ando et aL, 2001). However, a method to specifically modify gene expression in the juvenile or in the adult is still missing. Such a method would be a very powerful tool to understand gene function in differentiated tissues. We describe here an electroporation-based approach, which allows gene transfer in adult tissues. Its efficiency was assessed using a GFP (green fluorescent protein) dependent assay. We then used this method to disrupt the Fgf signalling pathway during the process of regeneration. (C) 2002 Wiley-Liss, Inc.