Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii

被引：189

作者：

Crozet, Pierre ^{[1
]}

Navarro, Francisco J. ^{[2
]}

Willmund, Felix ^{[3
]}

Mehrshahi, Payam ^{[2
]}

Bakowski, Kamil ^{[4
]}

Lauersen, Kyle J. ^{[5
]}

Perez-Perez, Maria-Esther ^{[6
]}

Auroy, Pascaline ^{[7
]}

Rovira, Aleix Gorchs ^{[2
]}

Sauret-Gueto, Susana ^{[2
]}

Niemeyer, Justus ^{[3
]}

Spaniol, Benjamin ^{[3
]}

Theis, Jasmine ^{[3
]}

Troesch, Raphael ^{[3
]}

Westrich, Lisa-Desiree ^{[3
]}

Vavitsas, Konstantinos ^{[4
,9
]}

Baier, Thomas ^{[5
]}

Huebner, Wolfgang ^{[8
]}

de Carpentier, Felix ^{[1
]}

Cassarini, Mathieu ^{[1
]}

Danon, Antoine ^{[1
]}

Henri, Julien ^{[1
]}

Marchand, Christophe H. ^{[1
]}

de Mia, Marcello ^{[1
]}

Sarkissian, Kevin ^{[1
]}

Baulcombe, David C. ^{[2
]}

Peltier, Gilles ^{[7
]}

Crespo, Jose-Luis ^{[6
]}

Kruse, Olaf ^{[5
]}

Jensen, Poul-Erik ^{[4
]}

Schroda, Michael ^{[3
]}

Smith, Alison G. ^{[2
]}

Lemaire, Stephane D. ^{[1
]}

机构：

[1] Sorbonne Univ, CNRS, UMR 8226, Inst Biol Physicochim, Paris, France

[2] Univ Cambridge, Dept Plant Sci, Cambridge CB2 3EA, England

[3] Tech Univ Kaiserslautern, Dept Biol, D-67663 Kaiserslautern, Germany

[4] Univ Copenhagen, Copenhagen Plant Sci Ctr, Dept Plant & Environm Sci, Copenhagen, Denmark

[5] Bielefeld Univ, Fac Biol, Ctr Biotechnol CeBiTec, D-33615 Bielefeld, Germany

[6] Univ Seville, CSIC, Inst Bioquim Vegetal & Fotosintesis, Seville 41092, Spain

[7] Aix Marseille Univ, Lab Bioenerget & Biotechnol Bacteries & Microalgu, CNRS, CEA,BIAM, St Paul Les Durance, France

[8] Bielefeld Univ, Dept Phys, Biomol Photon, D-33615 Bielefeld, Germany

[9] Univ Queensland, AIBN, Brisbane, Qld 4072, Australia

来源：

ACS SYNTHETIC BIOLOGY | 2018年 / 7卷 / 09期

基金：

英国生物技术与生命科学研究理事会;

关键词：

algal biotechnology; Chlamydomonas reinhardtii; modular cloning; synthetic biology; ZINC-FINGER NUCLEASES; HEAT-SHOCK FACTOR-1; GENE-EXPRESSION; INDUSTRIAL BIOTECHNOLOGY; EUKARYOTIC MICROALGAE; TRANSGENE EXPRESSION; COMBINATORIAL DESIGN; REVEALS; DNA; IDENTIFICATION;

D O I：

10.1021/acssynbio.8b00251

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Microalgae are regarded as promising organisms to develop innovative concepts based on their photosynthetic capacity that offers more sustainable production than heterotrophic hosts. However, to realize their potential as green cell factories, a major challenge is to make microalgae easier to engineer. A promising approach for rapid and predictable genetic manipulation is to use standardized synthetic biology tools and workflows. To this end we have developed a Modular Cloning toolkit for the green microalga Chlamydomonas reinhardtii. It is based on Golden Gate cloning with standard syntax, and comprises 119 openly distributed genetic parts, most of which have been functionally validated in several strains. It contains promoters, UTRs, terminators, tags, reporters, antibiotic resistance genes, and introns cloned in various positions to allow maximum modularity. The toolkit enables rapid building of engineered cells for both fundamental research and algal biotechnology. This work will make Chlamydomonas the next chassis for sustainable synthetic biology.

引用

页码：2074 / 2086

页数：25

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