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

AI Article Synopsis

  • * The microfluidic chip, made from PDMS and glass, features a flow-focusing microstructure and parallel microelectrodes, allowing for high efficiency without the need for costly electrical equipment.
  • * The transformation efficiency is significantly higher in the wall-less mutant (over three times) and the wild type (over two times) compared to traditional bulk phase electroporation, even with low DNA/cell ratios, which helps manage transgenic copy numbers for stable gene

Article 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.

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Source
http://dx.doi.org/10.1039/c2lc40360aDOI Listing

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