Linear double-stranded DNAs as innovative biological parts to implement genetic circuits in mammalian cells
Linear double-stranded DNAs as innovative biological parts to implement genetic circuits in mammalian cells
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线性双链 DNA 作为创新生物部件，在哺乳动物细胞中实现遗传电路
DOI：
10.1111/febs.14816
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发表时间：
2019-06-01
期刊：
影响因子：
5.4
通讯作者：
Zhang, Chunze
中科院分区：
文献类型：
--
作者：
Li, Shuai;Su, Weijun;Zhang, Chunze
合成生物学利用工程原理重新设计生物系统用于生物医学或工业目的。创新和应用原始生物部件构建基因电路将大大促进和加快合成生物学的发展。在这里，我们建立了两个或三个输入线性双链DNA（ldsDNA）为基础的布尔与门遗传电路在哺乳动物细胞。生物发光成像揭示了基于ldsDNA的布尔AND门电路在体内的可行性。DNA-PKcs是非同源末端连接的关键酶，抑制DNA-PKcs可显著减弱ldsDNA布尔与门电路的输出信号。我们进一步证明了带有额外末端随机核苷酸的ldsDNA可以通过布尔与门响应进行末端核苷酸缺失并产生框内蛋白。此外，具有相同重叠序列的ldsDNA或质粒也可以用作布尔与门遗传电路的输入信号。我们的工作建立ldsDNAs作为创新的生物部件，用于构建低噪声信号比布尔与门电路，在生物医学工程领域的应用潜力。
Synthetic biology employs engineering principles to redesign biological systems for biomedical or industrial purposes. Innovation and application of original biological parts for genetic circuit construction will significantly facilitate and expedite the development of synthetic biology. Here, we built two- or three-input linear double-stranded DNA (ldsDNA)-based Boolean AND gate genetic circuits in mammalian cells. Bioluminescence imaging revealed the feasibility of ldsDNA-based Boolean AND gate circuits in vivo. Inhibition of DNA-PKcs, a pivotal enzyme in nonhomologous end joining, significantly attenuated the output signals from ldsDNA-based Boolean AND gate circuits. We further showed that ldsDNA with additional terminal random nucleotide(s) could undergo end nucleotide deletion and generate in-frame proteins via the Boolean AND gate response. Additionally, ldsDNAs or plasmids with identical overlapping sequences could also serve as input signals for Boolean AND gate genetic circuits. Our work establishes ldsDNAs as innovative biological parts for building low noise-signal ratio Boolean AND gate circuits with application potential in biomedical engineering fields.