Two- and three-input TALE-based AND logic computation in embryonic stem cells.
Two- and three-input TALE-based AND logic computation in embryonic stem cells.
复制标题
DOI：
10.1093/nar/gkt758
复制
发表时间：
2013-11
影响因子：
14.9
通讯作者：
Silver PA
中科院分区：
文献类型：
--
作者：
Lienert F;Torella JP;Chen JH;Norsworthy M;Richardson RR;Silver PA
生物计算电路可以增强我们控制细胞功能的能力，在组织工程和医疗方面有潜在的应用。转录激活子样效应器(TALE)是合成基因调控电路中极具吸引力的组成部分，因为它们可以从头开始设计来靶向给定的DNA序列。我们在这里演示了TALES可以在哺乳动物细胞中进行布尔逻辑计算。使用分裂内含子蛋白质剪接策略，我们证明了功能TALE可以由两个不活跃的部分重组，从而产生两个输入和逻辑计算。我们进一步展示了哺乳动物细胞中的三段内含子剪接，并用它来进行三次输入和计算。使用随机和定向插入这些相对较大的遗传电路的方法，我们证明了基于TALL的逻辑电路在整合到小鼠胚胎干细胞的基因组中时是起作用的。比较相同基因组背景下的构建变体，我们调节了TALE反应启动子的强度，以提高这些电路的输出。我们的工作建立了分裂故事作为一种工具，用于建立逻辑计算，具有控制内源基因或转基因表达的潜力，以响应细胞信号的组合。
Biological computing circuits can enhance our ability to control cellular functions and have potential applications in tissue engineering and medical treatments. Transcriptional activator-like effectors (TALEs) represent attractive components of synthetic gene regulatory circuits, as they can be designed de novo to target a given DNA sequence. We here demonstrate that TALEs can perform Boolean logic computation in mammalian cells. Using a split-intein protein-splicing strategy, we show that a functional TALE can be reconstituted from two inactive parts, thus generating two-input AND logic computation. We further demonstrate three-piece intein splicing in mammalian cells and use it to perform three-input AND computation. Using methods for random as well as targeted insertion of these relatively large genetic circuits, we show that TALE-based logic circuits are functional when integrated into the genome of mouse embryonic stem cells. Comparing construct variants in the same genomic context, we modulated the strength of the TALE-responsive promoter to improve the output of these circuits. Our work establishes split TALEs as a tool for building logic computation with the potential of controlling expression of endogenous genes or transgenes in response to a combination of cellular signals.
登录
查看更多内容
影响因子：
4.2
作者：
Li, Juan;Sun, Wunchang;Liu, Xiang-Qin
通讯作者：
Liu, Xiang-Qin
影响因子：
56.9
作者：
Boch, Jens;Scholze, Heidi;Bonas, Ulla
通讯作者：
Bonas, Ulla
影响因子：
64.5
作者：
CHUNG, JH;WHITELEY, M;FELSENFELD, G
通讯作者：
FELSENFELD, G
影响因子：
10.5
作者：
Burrill, Devin R.;Inniss, Mara C.;Silver, Pamela A.
通讯作者：
Silver, Pamela A.
DOI：
10.1073/pnas.95.25.14628
发表时间：
1998-12-08
影响因子：
11.1
作者：
Beerli, RR;Segal, DJ;Barbas, CF
通讯作者：
Barbas, CF