High efficiency hydrodynamic bacterial electrotransformation
High efficiency hydrodynamic bacterial electrotransformation
复制标题
DOI：
10.1039/c6lc01309k
复制
发表时间：
2017-02-07
期刊：
影响因子：
6.1
通讯作者：
Buie, Cullen R.
中科院分区：
文献类型：
--
作者：
Garcia, Paulo A.;Ge, Zhifei;Buie, Cullen R.
合成生物学在解决人类和地球面临的紧迫挑战方面具有巨大潜力。利用合成生物学的全部潜力的一个技术挑战是对许多类型的细胞的低效率和低通量的遗传转化。本文讨论了一种新型微流体系统，用于提高细菌电转化效率和通量。我们的微流体系统是由微通道中的非均匀收缩组成的，以促进高电场和相对较小的施加电压来诱导电穿孔。此外，微流控装置具有低电场区域，以协助核酸进入细胞的电泳运输。该装置的特点是流体动力学控制的电场，允许细胞体验一个时间相关的电场，否则使用标准电子设备很难实现。结果表明，与传统的电穿孔比色皿相比，转化效率可提高约4倍，而通量可提高100-1000倍。这项工作将使微生物的高通量和高效率的遗传转化成为可能，促进基因工程生物的加速发展。
Synthetic biology holds great potential for addressing pressing challenges for mankind and our planet. One technical challenge in tapping into the full potential of synthetic biology is the low efficiency and low throughput of genetic transformation for many types of cells. In this paper, we discuss a novel microfluidic system for improving bacterial electrotransformation efficiency and throughput. Our microfluidic system is comprised of non-uniform constrictions in microchannels to facilitate high electric fields with relatively small applied voltages to induce electroporation. Additionally, the microfluidic device has regions of low electric field to assist in electrophoretic transport of nucleic acids into the cells. The device features hydrodynamically controlled electric fields that allow cells to experience a time dependent electric field that is otherwise difficult to achieve using standard electronics. Results suggest that transformation efficiency can be increased by similar to 4x, while throughput can increase by 100-1000x compared to traditional electroporation cuvettes. This work will enable high-throughput and high efficiency genetic transformation of microbes, facilitating accelerated development of genetically engineered organisms.