Droplet electroporation in microfluidics for efficient cell transformation with or without cell wall removal

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Title
Droplet electroporation in microfluidics for efficient cell transformation with or without cell wall removal
Author(s)
B Qu; Y J Eu; Won Joong Jeong; D P Kim
Bibliographic Citation
Lab on a Chip, vol. 12, no. 21, pp. 4483-4488
Publication Year
2012
Abstract
An efficient cell transformation method is presented that utilizes droplet electroporation on a microfluidic chip. Two types of green microalgae, a wall-less mutant and a wild type of Chlamydomonas reinhardtii, are used as model cells. The PDMS-glass electroporation chip is simply composed of a flow-focusing microstructure for generating cell-encapsulating droplets and a serpentine channel for better mixing of the content in the droplet, and five pairs of parallel microelectrodes on the glass slide, without involving any expensive electrical equipment. The transformation efficiency via the microfluidic electroporation is shown to be more than three orders of magnitude higher for the wall-less mutant, and more than two orders of magnitude higher for the wild type, which has its cell wall intact, than bulk phase electroporation under identical conditions. Furthermore, the microfluidic transformation is remarkably efficient even at a low DNA/cell ratio, facilitating ways of controlling the transgenic copy number, which is important for the stability of the transgene expression.
ISSN
1473-0197
Publisher
Royal Soc Chem
DOI
http://dx.doi.org/10.1039/c2lc40360a
Type
Article
Appears in Collections:
Synthetic Biology and Bioengineering Research Institute > Cell Factory Research Center > 1. Journal Articles
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