Efficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase.
Efficient deletion of LoxP-flanked selectable marker genes from the genome of transgenic pigs by an engineered Cre recombinase.
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DOI：
10.1007/s11248-020-00200-3
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发表时间：
2020-06
影响因子：
3
通讯作者：
Li Z
中科院分区：
文献类型：
--
作者：
Huang X;Zou X;Xu Z;Tang F;Shi J;Zheng E;Liu D;Moisyadi S;Urschitz J;Wu Z;Li Z
转基因（GM）猪为猪的遗传改良、人类健康和生命科学带来了巨大的希望。当生产转基因猪时，通常会将选择标记基因（SMG）引入其基因组中以供宿主细胞或动物识别。然而，转基因猪体内残留的 SMG 可能会产生多种副作用。为了避免 SMG 可能引起的副作用，应在商业化之前将其从转基因猪的基因组中去除。 Cre 重组酶通常用于从转基因动物基因组中删除位于 LoxP 位点侧翼的 SMG。尽管不含 SMG 的转基因猪是通过 Cre 介导的重组产生的，但更有效和更具成本效益的方法对于不含 SMG 的转基因猪的商业化至关重要。在本文中，我们描述了在一个构建体中含有细胞穿透和核定位信号肽的重组 Cre 蛋白的生产。这种工程化的 Cre 酶可以有效地切除从转基因猪中分离的培养成纤维细胞中的 LoxP 侧翼 SMG，然后将其用作核供体细胞，通过体细胞核移植产生带有所需转基因的活体无 SMG 转基因猪。这项研究描述了一种高效且成本低得多的生产不含 SMG 的转基因猪的方法。
Genetically modified (GM) pigs hold great promises for pig genetic improvement, human health and life science. When GM pigs are produced, selectable marker genes (SMGs) are usually introduced into their genomes for host cell or animal recognition. However, the SMGs that remain in GM pigs might have multiple side effects. To avoid the possible side effects caused by the SMGs, they should be removed from the genome of GM pigs before their commercialization. The Cre recombinase is commonly used to delete the LoxP sites-flanked SMGs from the genome of GM animals. Although SMG-free GM pigs have been generated by Cre-mediated recombination, more efficient and cost-effective approaches are essential for the commercialization of SMG-free GM pigs. In this article we describe the production of a recombinant Cre protein containing a cell-penetrating and a nuclear localization signal peptide in one construct. This engineered Cre enzyme can efficiently excise the LoxP-flanked SMGs in cultured fibroblasts isolated from a transgenic pig, which then can be used as nuclear donor cells to generate live SMG-free GM pigs harboring a desired transgene by somatic cell nuclear transfer. This study describes an efficient and far-less costly method for production of SMG-free GM pigs.
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影响因子：
4.6
作者：
Montag J;Petersen B;Flögel AK;Becker E;Lucas-Hahn A;Cost GJ;Mühlfeld C;Kraft T;Niemann H;Brenner B
通讯作者：
Brenner B
影响因子：
3.7
作者：
Zeng F;Li Z;Cai G;Gao W;Jiang G;Liu D;Urschitz J;Moisyadi S;Wu Z
通讯作者：
Wu Z
影响因子：
64.8
作者：
HAMMER, RE;PURSEL, VG;BRINSTER, RL
通讯作者：
BRINSTER, RL
影响因子：
3.6
作者：
Li, Zicong;Zeng, Fang;Wu, Zhenfang
通讯作者：
Wu, Zhenfang
影响因子：
--
作者：
McLellan, Micheal A;Rosenthal, Nadia A;Pinto, Alexander R
通讯作者：
Pinto, Alexander R