17 Mar Droplet microfluidics enables high-throughput CRISPR-based gene editing and screening
Abstract
“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 10 × 10 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 E. coli and (2) targeted engineering of the glutamine synthetase gene (glnA) and the blue-pigment synthetase gene (bpsA) to improve indigoidine production in E. coli.”
“a CRISPR-MAGE steps. The ones inside the box are performed on the chip. Cells are removed from the chip after the recovery step for induction and plating. b The microfluidic chip in a 3D printed holder (left), the electrode pattern (right), a top-view of an individual well, and a side-view schematic of a well. The chip is designed to contain 100 discrete reaction chambers with individually addressable electrodes for multiplexed CRISPR-MAGE recombineering, and its 384-well format design can be interfaced with lab automation equipment. c Droplets containing plasmids and cells are dispensed into each chamber through the inlet port, mixed by electrowetting, and electroporated by applying a voltage pulse.” Reproduced under Creative Commons Attribution 4.0 International License from Iwai, K., Wehrs, M., Garber, M. et al. Scalable and automated CRISPR-based strain engineering using droplet microfluidics. Microsyst Nanoeng 8, 31 (2022)..
Figures and the abstract are reproduced from Iwai, K., Wehrs, M., Garber, M. et al. Scalable and automated CRISPR-based strain engineering using droplet microfluidics. Microsyst Nanoeng 8, 31 (2022). https://doi.org/10.1038/s41378-022-00357-3 under Creative Commons Attribution 4.0 International License
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