Identification and re-addressing of a transcriptionally permissive locus in the porcine genome
Identification and re-addressing of a transcriptionally permissive locus in the porcine genome
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猪基因组中转录允许位点的鉴定和重新寻址
DOI：
10.1007/s11248-015-9914-4
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发表时间：
2016
影响因子：
3
通讯作者：
Kues WA
中科院分区：
文献类型：
--
作者：
Garrels W;Mukherjee A;Holler S;Cleve N;Talluri TR;Barg-Kues B;Diederich M;Köhler P;Diederich M;Petersen B;Lucas-Hahn;Niemann H;Izsvac Z;Ivics Z;Kues WA
最近，我们建立了睡美人转座子系统，用于猪的生殖系感受态转基因。在这里，我们扩展了这种方法，重新定位转座子标记的位点的位点特异性基因敲入，并产生了一个同基因组的小猪携带原始转座子或重新定位的事件。进行Cre-loxP介导的标签转座子与不同报告基因的盒交换，然后进行流式细胞术分选和重组细胞的体细胞核转移。平行地，将原始细胞用于体细胞核转移以产生克隆同胞，从而产生在相同染色体位置携带不同报告转座子的仔猪的克隆群组。重要的是，这种策略取代了对抗生素选择标记的需要。这种方法扩大了家畜基因组工程技术的武库，并将促进人类疾病的大型动物模型的开发。潜在地，猪的同基因组将有助于在新型细胞疗法的临床前评估中对移植细胞的重要跟踪。
Recently, we established theSleeping Beautytransposon system for germ line competent transgenesis in the pig. Here, we extend this approach to re-target a transposon-tagged locus for a site-specific gene knock-in, and generated a syngeneic cohort of piglets carrying either the original transposon or the re-targeted event. A Cre-loxP-mediated cassette exchange of the tagging transposon with a different reporter gene was performed, followed by flow cytometric sorting and somatic cell nuclear transfer of recombined cells. In parallel, the original cells were employed in somatic cell nuclear transfer to generate clone siblings, thereby resulting in a clone cohort of piglets carrying different reporter transposons at an identical chromosomal location. Importantly, this strategy supersedes the need for an antibiotic selection marker. This approach expands the arsenal of genome engineering technologies in domestic animals, and will facilitate the development of large animal models for human diseases. Potentially, the syngeneic cohort of pigs will be instrumental for vital tracking of transplanted cells in pre-clinical assessments of novel cell therapies.
影响因子：
4.8
作者：
Esteban, Miguel A.;Xu, Jianyong;Pei, Duanqing
通讯作者：
Pei, Duanqing
DOI：
10.1073/pnas.1106422108
发表时间：
2011-07-19
影响因子：
11.1
作者：
Hauschild, Janet;Petersen, Bjoern;Niemann, Heiner
通讯作者：
Niemann, Heiner
影响因子：
0.9
作者：
Shigeta, T.;Hsu, H. -C.;Nagashima, H.
通讯作者：
Nagashima, H.