Protecting a transgene expression from the HAC-based vector by different chromatin insulators.
Protecting a transgene expression from the HAC-based vector by different chromatin insulators.
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DOI：
10.1007/s00018-013-1362-9
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发表时间：
2013-10
影响因子：
8
通讯作者：
Kouprina, Natalay
中科院分区：
文献类型：
--
作者：
Lee, Nicholas Co;Kononenko, Artem V.;Lee, Hee-Sheung;Tolkunova, Elena N.;Liskovykh, Mikhail A.;Masumoto, Hiroshi;Earnshaw, William C.;Tomilin, Alexey N.;Larionov, Vladimir;Kouprina, Natalay
人类人工染色体（HACs）是一种载体，它为基因的传递和表达提供了容量和稳定性的优势。一些研究甚至证明了它们在基因缺陷受体细胞系和动物转基因中的基因互补作用。最近，我们构建了一个先进的基于BAC的载体，alphoidtetO-HAC，与条件着丝粒。在此HAC中，基因加载位点插入到着丝粒组装和维护的中心染色质结构域的关键。虽然根据定义，该结构域是允许转录的，但还没有关于中心染色质内转基因表达的长期研究。在这项研究中，我们比较了三个染色质绝缘体，cHS 4，γ-卫星DNA，和tDNA，对插入到alphoidtetO-HAC载体的EGFP转基因的表达的影响。绝缘子的功能是必不可少的稳定表达的转基因在中央染色质。在两个分析的宿主细胞系中，由两个功能性tRNA基因拷贝组成的tRNA绝缘子显示出最高的屏障活性。我们推断，接近中央染色质不能保护缺乏染色质绝缘子的基因免受表观遗传沉默。因此，防止中心染色质中基因沉默的屏障元件将有助于使用HAC载体优化转基因。本文的在线版本（doi：10.1007/s 00018 -013-1362-9）包含补充材料，可供授权用户使用。
Human artificial chromosomes (HACs) are vectors that offer advantages of capacity and stability for gene delivery and expression. Several studies have even demonstrated their use for gene complementation in gene-deficient recipient cell lines and animal transgenesis. Recently, we constructed an advance HAC-based vector, alphoidtetO-HAC, with a conditional centromere. In this HAC, a gene-loading site was inserted into a centrochromatin domain critical for kinetochore assembly and maintenance. While by definition this domain is permissive for transcription, there have been no long-term studies on transgene expression within centrochromatin. In this study, we compared the effects of three chromatin insulators, cHS4, gamma-satellite DNA, and tDNA, on the expression of an EGFP transgene inserted into the alphoidtetO-HAC vector. Insulator function was essential for stable expression of the transgene in centrochromatin. In two analyzed host cell lines, a tDNA insulator composed of two functional copies of tRNA genes showed the highest barrier activity. We infer that proximity to centrochromatin does not protect genes lacking chromatin insulators from epigenetic silencing. Barrier elements that prevent gene silencing in centrochromatin would thus help to optimize transgenesis using HAC vectors. The online version of this article (doi:10.1007/s00018-013-1362-9) contains supplementary material, which is available to authorized users.
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影响因子：
11.4
作者：
Ohzeki, Jun-ichirou;Bergmann, Jan H.;Kouprina, Natalay;Noskov, Vladimir N.;Nakano, Megumi;Kimura, Hiroshi;Earnshaw, William C.;Larionov, Vladimir;Masumoto, Hiroshi
通讯作者：
Masumoto, Hiroshi
影响因子：
7
作者：
Kim, Jung-Hyun;Ebersole, Thomas;Larionov, Vladimir
通讯作者：
Larionov, Vladimir
影响因子：
14.8
作者：
Kouprina, Natalay;Larionov, Vladimir
通讯作者：
Larionov, Vladimir
影响因子：
4.3
作者：
Ebersole, Thomas;Kim, Jung-Hyun;Larionov, Vladimir
通讯作者：
Larionov, Vladimir
DOI：
10.1007/978-1-61779-099-7_7
发表时间：
2011-01-01
期刊：
MAMMALIAN CHROMOSOME ENGINEERING: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ikeno, Masashi;Suzuki, Nobutaka
通讯作者：
Suzuki, Nobutaka