Testing a three-finger zinc finger nuclease using a GFP reporter system.
Testing a three-finger zinc finger nuclease using a GFP reporter system.
复制标题
使用 GFP 报告系统测试三指锌指核酸酶。
DOI：
10.1101/pdb.prot5531
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发表时间：
2010
影响因子：
--
通讯作者：
Porteus,Matthew
中科院分区：
文献类型：
--
作者：
Porteus,Matthew
Homologous recombination is the most precise way to manipulate the genome. It has been used extensively in bacteria, yeast, murine embryonic stem cells, and a few other specialized cell lines, but it has not been available in other systems such as mammalian somatic cells. However, the creation of a gene-specific DNA double-strand break can stimulate homologous recombination by several-thousandfold in mammalian somatic cells. These double-strand breaks can be created in mammalian genomes by zinc finger nucleases (ZFNs), artificial proteins in which a zinc finger DNA-binding domain is fused to a nonspecific nuclease domain. This protocol describes how to test newly designed ZFNs using a cell-based green fluorescent protein (GFP) reporter assay to determine if they are active in a mammalian cell-culture-based system.
