Elsevier

Algal Research

Volume 33, July 2018, Pages 448-455
Algal Research

Improved DNA/protein delivery in microalgae – A simple and reliable method for the prediction of optimal electroporation settings

https://doi.org/10.1016/j.algal.2018.06.021Get rights and content
Under a Creative Commons license
open access

Highlights

  • Electroporation settings were successfully predicted for four microalgae species.

  • DNA and proteins were delivered into the cells to evaluate predicted settings.

  • High transformation efficiencies were accomplished in A. obliquus.

  • Low cell concentration, light intensity and fragment size increase efficiencies.

Abstract

Genetic transformation of microalgae remains a challenge due to poor intracellular delivery of exogenous molecules. This limitation is caused by the structure and composition of the cell wall and cell membrane of each species. Moreover, successful delivery of proteins or nucleic acids cannot be assessed by determining transformability since their functionality is not always known in the studied microorganism. We propose a quick and effective screening tool for the prediction and optimization of electroporation settings by monitoring cell permeability and viability using Sytox Green and propidium iodide respectively. We determined voltage settings for the microalgae Chlamydomonas reinhardtii, Chlorella vulgaris, Neochloris oleoabundans and Acutodesmus obliquus. To evaluate the predicted settings, we delivered labelled DNA and proteins into the cells. We demonstrated that high transformation efficiencies can be accomplished when predicted values were applied with functional plasmids. Additionally, we increased transformation efficiencies by testing cell concentrations, light intensities and fragment sizes. This method can be used to determine suitable transformation conditions for non-transformed microalgae species and to increase the insight on established transformation protocols.

Keywords

Electroporation
Green microalgae
Labelled DNA
Labelled protein
Sytox Green
Propidium iodide

Cited by (0)

1

Both authors contributed equally to this work.