Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii
Abstract 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 induc...
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doaj-b31009f1deb74297a8247bd9427711802020-11-24T23:14:53ZengBMCBiotechnology for Biofuels1754-68342017-11-011011910.1186/s13068-017-0941-7Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtiiYuting Wang0Xinqin Jiang1Changxing Hu2Ting Sun3Zhiyong Zeng4Xiaoqi Cai5Hui Li6Zhangli Hu7Guangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityGuangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Key Laboratory of Plant Epigenetics, College of Life Sciences and Oceanography, Shenzhen UniversityAbstract 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.http://link.springer.com/article/10.1186/s13068-017-0941-7OptogeneticLight-inducible systemMicroRNABio-hydrogen productionMicroalgaChlamydomonas reinhardtii |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yuting Wang Xinqin Jiang Changxing Hu Ting Sun Zhiyong Zeng Xiaoqi Cai Hui Li Zhangli Hu |
spellingShingle |
Yuting Wang Xinqin Jiang Changxing Hu Ting Sun Zhiyong Zeng Xiaoqi Cai Hui Li Zhangli Hu Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii Biotechnology for Biofuels Optogenetic Light-inducible system MicroRNA Bio-hydrogen production Microalga Chlamydomonas reinhardtii |
author_facet |
Yuting Wang Xinqin Jiang Changxing Hu Ting Sun Zhiyong Zeng Xiaoqi Cai Hui Li Zhangli Hu |
author_sort |
Yuting Wang |
title |
Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii |
title_short |
Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii |
title_full |
Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii |
title_fullStr |
Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii |
title_full_unstemmed |
Optogenetic regulation of artificial microRNA improves H2 production in green alga Chlamydomonas reinhardtii |
title_sort |
optogenetic regulation of artificial microrna improves h2 production in green alga chlamydomonas reinhardtii |
publisher |
BMC |
series |
Biotechnology for Biofuels |
issn |
1754-6834 |
publishDate |
2017-11-01 |
description |
Abstract 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. |
topic |
Optogenetic Light-inducible system MicroRNA Bio-hydrogen production Microalga Chlamydomonas reinhardtii |
url |
http://link.springer.com/article/10.1186/s13068-017-0941-7 |
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