Generation of medaka gene knockout models by target-selected mutagenesis.
Generation of medaka gene knockout models by target-selected mutagenesis.
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DOI：
10.1186/gb-2006-7-12-r116
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发表时间：
2006
期刊：
影响因子：
12.3
通讯作者：
Cuppen E
中科院分区：
文献类型：
--
作者：
Taniguchi Y;Takeda S;Furutani-Seiki M;Kamei Y;Todo T;Sasado T;Deguchi T;Kondoh H;Mudde J;Yamazoe M;Hidaka M;Mitani H;Toyoda A;Sakaki Y;Plasterk RH;Cuppen E
一个反向遗传学方法的常规一代青（青）基因敲除的描述和应用，以创建一个冷冻保存的资源含有大多数青基因敲除。我们建立了一个反向遗传学的方法，常规一代青鳉（Oryzias latipes）基因敲除。利用高通量重测序技术对ENU诱变鱼的冻存文库进行了点突变筛选。无义和剪接位点突变检索Blm，Sirt1，帕金和p53基因和p53突变体的功能表征表明p53功能的完全敲除。目前冷冻保存的资源预计将包含大多数青鳉基因的敲除。
A reverse genetics approach for the routine generation of medaka (Oryzias latipes) gene knockouts is described and applied to create a cryopreserved resource containing knockouts for most medaka genes. We have established a reverse genetics approach for the routine generation of medaka (Oryzias latipes) gene knockouts. A cryopreserved library of N-ethyl-N-nitrosourea (ENU) mutagenized fish was screened by high-throughput resequencing for induced point mutations. Nonsense and splice site mutations were retrieved for the Blm, Sirt1, Parkin and p53 genes and functional characterization of p53 mutants indicated a complete knockout of p53 function. The current cryopreserved resource is expected to contain knockouts for most medaka genes.