Intronically encoded siRNAs improve dynamic range of mammalian gene regulation systems and toggle switch.
Intronically encoded siRNAs improve dynamic range of mammalian gene regulation systems and toggle switch.
复制标题
固有编码的siRNA改善了哺乳动物基因调节系统和拨动开关的动态范围。
DOI：
10.1093/nar/gkn443
复制
发表时间：
2008-09
影响因子：
14.9
通讯作者：
Fussenegger, Martin
中科院分区：
文献类型：
--
作者：
Greber, David;El-Baba, Marie Daoud;Fussenegger, Martin
条件基因表达的应用，无论是用于治疗还是基础研究目的，都越来越需要表现出更严格控制特性的哺乳动物基因控制系统。虽然已经使用了许多方法来改善广泛使用的Tet-regulatory系统，但许多应用，特别是关于合成基因网络的工程，将需要更广泛的严格执行的基因控制系统。在这里，一般适用的方法，利用内含子编码的siRNA的相关transregulator构建体，和siRNA序列特异性标签的报告构建体，以尽量减少基础基因的活性在关闭状态下的一系列常见的基因控制系统。为了证明对残余表达的严格控制，该方法成功地用于条件性表达毒性蛋白RipDD和Linamarase。内含子siRNA概念也被扩展以创建新一代紧凑的、单载体的、自诱导的siRNA载体。最后，使用改进的调节系统，对哺乳动物表观遗传拨动开关进行工程改造，与等效的非内含子系统相比，其在小鼠中表现出上级体外和体内诱导特征。
Applications of conditional gene expression, whether for therapeutic or basic research purposes, are increasingly requiring mammalian gene control systems that exhibit far tighter control properties. While numerous approaches have been used to improve the widely used Tet-regulatory system, many applications, particularly with respect to the engineering of synthetic gene networks, will require a broader range of tightly performing gene control systems. Here, a generically applicable approach is described that utilizes intronically encoded siRNA on the relevant transregulator construct, and siRNA sequence-specific tags on the reporter construct, to minimize basal gene activity in the off-state of a range of common gene control systems. To demonstrate tight control of residual expression the approach was successfully used to conditionally express the toxic proteins RipDD and Linamarase. The intronic siRNA concept was also extended to create a new generation of compact, single-vector, autoinducible siRNA vectors. Finally, using improved regulation systems a mammalian epigenetic toggle switch was engineered that exhibited superior in vitro and in vivo induction characteristics in mice compared to the equivalent non-intronic system.
登录
查看更多内容
DOI：
10.1073/pnas.0500345102
发表时间：
2005-07-05
影响因子：
11.1
作者：
Kramer, BP;Fussenegger, M
通讯作者：
Fussenegger, M
影响因子：
3.5
作者：
Aubrecht, J;Manivasakam, P;Schiestl, RH
通讯作者：
Schiestl, RH
影响因子：
14.9
作者：
Baron, U;Gossen, M;Bujard, H
通讯作者：
Bujard, H
DOI：
10.1073/pnas.90.5.1657
发表时间：
1993-03-01
影响因子：
11.1
作者：
BRASELMANN, S;GRANINGER, P;BUSSLINGER, M
通讯作者：
BUSSLINGER, M
影响因子：
46.9
作者：
Fussenegger, M;Morris, RP;Bailey, JE
通讯作者：
Bailey, JE