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Biology 2018, 7(4), 46; https://doi.org/10.3390/biology7040046

Selectable Markers and Reporter Genes for Engineering the Chloroplast of Chlamydomonas reinhardtii

1
Department of Life Sciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK
2
School of Biological Sciences and Engineering, Yachay-Tech University, Hacienda San José, Urcuquí-Imbabura 100650, Ecuador
3
Institute of Structural & Molecular Biology, University College London, Gower Street, London WC1E 6BT, UK
*
Author to whom correspondence should be addressed.
Received: 10 September 2018 / Revised: 2 October 2018 / Accepted: 3 October 2018 / Published: 10 October 2018
(This article belongs to the Special Issue Microalgal Biotechnology)
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Abstract

Chlamydomonas reinhardtii is a model alga of increasing interest as a cell factory for the production of valuable compounds, including therapeutic proteins and bioactive metabolites. Expression of foreign genes in the chloroplast is particularly advantageous as: (i) accumulation of product in this sub-cellular compartment minimises potential toxicity to the rest of the cell; (ii) genes can integrate at specific loci of the chloroplast genome (plastome) by homologous recombination; (iii) the high ploidy of the plastome and the high-level expression of chloroplast genes can be exploited to achieve levels of recombinant protein as high as 5% total cell protein; (iv) the lack of any gene silencing mechanisms in the chloroplast ensures stable expression of transgenes. However, the generation of C. reinhardtii chloroplast transformants requires efficient methods of selection, and ideally methods for subsequent marker removal. Additionally, the use of reporter genes is critical to achieving a comprehensive understanding of gene expression, thereby informing experimental design for recombinant applications. This review discusses currently available selection and reporter systems for chloroplast engineering in C. reinhardtii, as well as those used for chloroplast engineering in higher plants and other microalgae, and looks to the future in terms of possible new markers and reporters that will further advance the C. reinhardtii chloroplast as an expression platform. View Full-Text
Keywords: Chlamydomonas reinhardtii; chloroplast engineering; selectable markers; reporter genes Chlamydomonas reinhardtii; chloroplast engineering; selectable markers; reporter genes
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Esland, L.; Larrea-Alvarez, M.; Purton, S. Selectable Markers and Reporter Genes for Engineering the Chloroplast of Chlamydomonas reinhardtii. Biology 2018, 7, 46.

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