We present a droplet-based microfluidic system that enables CRISPR-based gene editing and high-throughput screening on a chip.The microfluidic device contains a 10x10 element array,and each element contains sets of el...We present a droplet-based microfluidic system that enables CRISPR-based gene editing and high-throughput screening on a chip.The microfluidic device contains a 10x10 element array,and each element contains sets of electrodes for two electric field-actuated operations:electrowetting for merging droplets to mix reagents and electroporation for transformation.This device can perform up to 100 genetic modification reactions in parallel,providing a scalable platform for generating the large number of engineered strains required for the combinatorial optimization of genetic pathways and predictable bioengineering.We demonstrate the system's capabilities through the CRISPR-based engineering of two test cases:(1)disruption of the function of the enzyme galactokinase(galK)in£coli and(2)targeted engineering of the glutamine synthetase gene(glnA)and the blue-pigment synthetase gene(bpsA)to improve indigoidine production in£coli.展开更多
基金supported by the Basque Government through the BERC 2014-2017 program and the Spanish Ministry of Economy and Competitiveness MINECO through the BCAM Severo Ochoa excellence accreditation SEV-2013-0323Sandia National Laboratories is a multimission laboratory managed and operated by National Technology and Engineering Solutions of Sandia,LLC,a wholly owned subsidiary of Honeywell International,Inc.,for the U.S.Department of Energy's National Nuclear Security Administration under contract DE-NA-0003525.
文摘We present a droplet-based microfluidic system that enables CRISPR-based gene editing and high-throughput screening on a chip.The microfluidic device contains a 10x10 element array,and each element contains sets of electrodes for two electric field-actuated operations:electrowetting for merging droplets to mix reagents and electroporation for transformation.This device can perform up to 100 genetic modification reactions in parallel,providing a scalable platform for generating the large number of engineered strains required for the combinatorial optimization of genetic pathways and predictable bioengineering.We demonstrate the system's capabilities through the CRISPR-based engineering of two test cases:(1)disruption of the function of the enzyme galactokinase(galK)in£coli and(2)targeted engineering of the glutamine synthetase gene(glnA)and the blue-pigment synthetase gene(bpsA)to improve indigoidine production in£coli.
