Site-specific recombination in the chicken genome using Flipase recombinase-mediated cassette exchange
Site-specific recombination in the chicken genome using Flipase recombinase-mediated cassette exchange
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DOI：
10.1096/fj.15-274712
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发表时间：
2016-02-01
期刊：
影响因子：
4.8
通讯作者：
Han, Jae Yong
中科院分区：
文献类型：
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作者：
Lee, Hong Jo;Lee, Hyung Chul;Han, Jae Yong
靶向基因组重组已应用于不同的研究领域，并具有广泛的应用前景。特别是，基因组中支持整合基因的稳健和普遍表达的特定基因座的发现以及基因组编辑技术的发展促进了各个科学领域的快速发展。在这项研究中，我们生产的转基因（TG）鸡，可以诱导重组酶介导的基因盒交换（RMCE），位点特异性重组技术之一，并确认RMCE在TG鸡源性细胞。因此，我们建立了TG鸡品系，其在鸡基因组中具有由piggyBac转座介导的Flipase（Flp）识别靶标（FRT）对。转基因整合模式在转基因鸡各品系中存在差异，这种整合的多样性导致外源基因在转基因鸡各组织中表达水平的差异。此外，替换的基因盒成功表达，并保持在TG鸡源细胞的FRT优势位点的RMCE。这些结果表明，有针对性的基因组重组技术与RMCE可以适应TG鸡模型，该技术将适用于特定的基因调控，通过顺式元件插入和定制的功能蛋白表达在预测的水平，没有表观遗传的影响。
Targeted genome recombination has been applied in diverse research fields and has a wide range of possible applications. In particular, the discovery of specific loci in the genome that support robust and ubiquitous expression of integrated genes and the development of genome-editing technology have facilitated rapid advances in various scientific areas. In this study, we produced transgenic (TG) chickens that can induce recombinase-mediated gene cassette exchange (RMCE), one of the site-specific recombination technologies, and confirmed RMCE in TG chicken-derived cells. As a result, we established TG chicken lines that have, Flipase (Flp) recognition target (FRT) pairs in the chicken genome, mediated by piggyBac transposition. The transgene integration patterns were diverse in each TG chicken line, and the integration diversity resulted in diverse levels of expression of exogenous genes in each tissue of the TG chickens. In addition, the replaced gene cassette was expressed successfully and maintained by RMCE in the FRT predominant loci of TGchicken-derived cells. These results indicate that targeted genome recombination technology with RMCE could be adaptable to TG chicken models and that the technology would be applicable to specific gene regulation by cis-element insertion and customized expression of functional proteins at predicted levels without epigenetic influence.