Characterization of gene expression regulation using D-RECS polymer by enzymatic reaction for an effective design of enzyme-responsive gene regulator.
Characterization of gene expression regulation using D-RECS polymer by enzymatic reaction for an effective design of enzyme-responsive gene regulator.
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使用 D-RECS 聚合物通过酶反应表征基因表达调控，以有效设计酶响应基因调控因子。
DOI：
10.1016/j.jconrel.2010.01.013
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Y. Katayama
中科院分区：
文献类型：
--
作者：
Yuko T. Sato;K. Kawamura;T. Niidome;Y. Katayama
靶向基因递送系统的发展引起了人们的广泛关注。然而，靶细胞特异性基因递送尚未实现。然而，如果基因表达可以用基因载体调节，这种方法可能是靶向特异性基因治疗的有力工具。我们以前曾报道过一些新的人工基因调控系统响应细胞内信号酶通过使用接枝型聚合物，含有底物肽（共轭物）的目标酶。所述缀合物能够在无细胞系统、培养的细胞以及体内系统中调节基因转录。我们将这一概念称为“D-RECS”，意思是响应细胞信号的药物或基因递送系统。在本研究中，为了阐明通过酶促反应使用缀合物的基因表达调控的机制，我们表征了缀合物/DNA复合物的高级结构和使用Caspase-3响应系统作为典型实例的这种缀合物控制基因表达的能力。此外，我们比较了这些特性与底物肽/DNA复合物。因此，我们阐明了聚合物骨架对基因调控的重要性。因为我们的缀合物含有大量的中性聚合物链，所以用聚合物链包裹DNA链起到抑制序列识别酶如RNA聚合酶或限制酶的“滑动”或“跳跃”的作用，使得缀合物可以有效地抑制基因表达或位点特异性切割。此外，我们的缀合物与DNA形成松散堆积的复合物。这种行为对于基因转录的酶促调节是必不可少的，因为复合物允许必需酶如胱天蛋白酶-3的进入，但禁止RNA聚合酶的进入和/或滑动。
The development of targeted gene delivery systems has attracted much attention. Nevertheless, target cell-specific gene delivery has not been realized. However, if gene expression can be regulated with a gene carrier, this method could be a powerful tool for target specific gene therapy. We have previously reported some novel artificial gene-regulation systems responding to intracellular signaling enzymes by using graft-type polymers that contain substrate peptide (conjugate) for a target enzyme. The conjugate is able to regulate gene transcription in a cell-free system, in cultivated cells and also in in vivo system. We termed this concept ‘D-RECS' which means Drug or gene delivery system responding to cellular signals. In the present study, in order to elucidate the mechanism of this gene expression regulation using the conjugate through enzymatic reactions, we characterized a high order structure of the conjugate/DNA complex and the ability of this conjugate to control gene expression using a Caspase-3 responsive system as a typical example. In addition, we compared these properties with those of a substrate peptide/DNA complex. As a result, we elucidated the importance of the polymer backbone for gene regulation. Because our conjugate contains a large amount of neutral polymer chain, wrapping of the DNA strand with a polymer chain acts as a suppressor for ‘sliding’ or ‘jumping’ of sequence-recognizing enzymes such as RNA polymerase or restriction enzymes so that the conjugate can efficiently suppress gene expression or site-specific cleavage. Furthermore, our conjugates formed a loosely packed complex with DNA. This behavior is essential for enzymatic regulation of gene transcription because the complex permits access of essential enzymes such as Caspase-3 but prohibits the access and/or sliding of RNA polymerase.
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影响因子：
6.2
作者：
K. Kawamura;Jun Oishi;Jeong-Hun Kang;K. Kodama;T. Sonoda;M. Murata;T. Niidome;Y. Katayama
通讯作者：
K. Kawamura;Jun Oishi;Jeong-Hun Kang;K. Kodama;T. Sonoda;M. Murata;T. Niidome;Y. Katayama
影响因子：
64.8
作者：
Sano, Masanori;Minamino, Tohru;Komuro, Issei
通讯作者：
Komuro, Issei
单分子观察揭示质粒 DNA 折叠转变为松散折叠状态
DOI：
--
发表时间：
2005
期刊：
FEBS Lett. 579(14)
影响因子：
--
作者：
Sato;Y.T.et al.
通讯作者：
Y.T.et al.
影响因子：
15
作者：
Kang, Jeong-Hun;Asai, Daisuke;Katayama, Yoshiki
通讯作者：
Katayama, Yoshiki
影响因子：
6.2
作者：
Akitaya, Tatsuo;Seno, Asako;Yoshikawa, Kenichi
通讯作者：
Yoshikawa, Kenichi