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Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii

BACKGROUND: Chlamydomonas reinhardtii is an ideal model organism not only for the study of basic metabolic processes in both plants and animals but also the production of biofuels including hydrogen. Transgenic analysis of C. reinhardtii is now well established and very convenient, but inducible exo...

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Autores principales: Wang, Yuting, Jiang, Xinqin, Hu, Changxing, Sun, Ting, Zeng, Zhiyong, Cai, Xiaoqi, Li, Hui, Hu, Zhangli
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5678773/
https://www.ncbi.nlm.nih.gov/pubmed/29151886
http://dx.doi.org/10.1186/s13068-017-0941-7
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author Wang, Yuting
Jiang, Xinqin
Hu, Changxing
Sun, Ting
Zeng, Zhiyong
Cai, Xiaoqi
Li, Hui
Hu, Zhangli
author_facet Wang, Yuting
Jiang, Xinqin
Hu, Changxing
Sun, Ting
Zeng, Zhiyong
Cai, Xiaoqi
Li, Hui
Hu, Zhangli
author_sort Wang, Yuting
collection PubMed
description BACKGROUND: Chlamydomonas reinhardtii is an ideal model organism not only for the study of basic metabolic processes in both plants and animals but also the production of biofuels including hydrogen. Transgenic analysis of C. reinhardtii is now well established and very convenient, but inducible exogenous gene expression systems remain under-studied. The most commonly used heat shock-inducible system has serious effects on algal cell growth and is difficult and costly to control in large-scale culture. Previous studies of hydrogen photoproduction in Chlamydomonas also use this heat-inducible system to activate target gene transcription and hydrogen synthesis. RESULTS: Here we describe a blue light-inducible system with which we achieved optogenetic regulation of target gene expression in C. reinhardtii. This light-inducible system was engineered in a photosynthetic organism for the first time. The photo-inducible heterodimerizing proteins CRY2 and CIB1 were fused to VP16 transcription activation domain and the GAL4 DNA-binding domain, respectively. This scheme allows for transcription activation of the target gene downstream of the activation sequence in response to blue light. Using this system, we successfully engineered blue light-inducible hydrogen-producing transgenic alga. The transgenic alga was cultured under red light and grew approximately normally until logarithmic phase. When illuminated with blue light, the transgenic alga expressed the artificial miRNA targeting photosynthetic system D1 protein, and altered hydrogen production was observed. CONCLUSIONS: The light-inducible system successfully activated the artificial miRNA and, consequently, regulation of its target gene under blue light. Moreover, hydrogen production was enhanced using this system, indicating a more convenient and efficient approach for gene expression regulation in large-scale microalgae cultivation. This optogenetic gene control system is a useful tool for gene regulation and also establishes a novel way to improve hydrogen production in green algae. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-017-0941-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-56787732017-11-17 Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii Wang, Yuting Jiang, Xinqin Hu, Changxing Sun, Ting Zeng, Zhiyong Cai, Xiaoqi Li, Hui Hu, Zhangli Biotechnol Biofuels Research BACKGROUND: Chlamydomonas reinhardtii is an ideal model organism not only for the study of basic metabolic processes in both plants and animals but also the production of biofuels including hydrogen. Transgenic analysis of C. reinhardtii is now well established and very convenient, but inducible exogenous gene expression systems remain under-studied. The most commonly used heat shock-inducible system has serious effects on algal cell growth and is difficult and costly to control in large-scale culture. Previous studies of hydrogen photoproduction in Chlamydomonas also use this heat-inducible system to activate target gene transcription and hydrogen synthesis. RESULTS: Here we describe a blue light-inducible system with which we achieved optogenetic regulation of target gene expression in C. reinhardtii. This light-inducible system was engineered in a photosynthetic organism for the first time. The photo-inducible heterodimerizing proteins CRY2 and CIB1 were fused to VP16 transcription activation domain and the GAL4 DNA-binding domain, respectively. This scheme allows for transcription activation of the target gene downstream of the activation sequence in response to blue light. Using this system, we successfully engineered blue light-inducible hydrogen-producing transgenic alga. The transgenic alga was cultured under red light and grew approximately normally until logarithmic phase. When illuminated with blue light, the transgenic alga expressed the artificial miRNA targeting photosynthetic system D1 protein, and altered hydrogen production was observed. CONCLUSIONS: The light-inducible system successfully activated the artificial miRNA and, consequently, regulation of its target gene under blue light. Moreover, hydrogen production was enhanced using this system, indicating a more convenient and efficient approach for gene expression regulation in large-scale microalgae cultivation. This optogenetic gene control system is a useful tool for gene regulation and also establishes a novel way to improve hydrogen production in green algae. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13068-017-0941-7) contains supplementary material, which is available to authorized users. BioMed Central 2017-11-07 /pmc/articles/PMC5678773/ /pubmed/29151886 http://dx.doi.org/10.1186/s13068-017-0941-7 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Wang, Yuting
Jiang, Xinqin
Hu, Changxing
Sun, Ting
Zeng, Zhiyong
Cai, Xiaoqi
Li, Hui
Hu, Zhangli
Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii
title Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii
title_full Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii
title_fullStr Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii
title_full_unstemmed Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii
title_short Optogenetic regulation of artificial microRNA improves H(2) production in green alga Chlamydomonas reinhardtii
title_sort optogenetic regulation of artificial microrna improves h(2) production in green alga chlamydomonas reinhardtii
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5678773/
https://www.ncbi.nlm.nih.gov/pubmed/29151886
http://dx.doi.org/10.1186/s13068-017-0941-7
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