Endoribonuclease-Based Two-Component Repressor Systems for Tight Gene Expression Control in Plants

Liang, Yan; Richardson, Sarah; Yan, Jingwei; Benites, Veronica T.; Cheng-Yue, Clarabelle; Tran, Thu; Mortimer, Jenny; Mukhopadhyay, Aindrila; Keasling, Jay D.; Scheller, Henrik V.; Loqué, Dominique

doi:10.1021/acssynbio.6b00295

Title: Endoribonuclease-Based Two-Component Repressor Systems for Tight Gene Expression Control in Plants

Journal Article · 17 January 2017 · ACS Synthetic Biology

DOI: https://doi.org/10.1021/acssynbio.6b00295 · OSTI ID:1361519

Liang, Yan ^[1];

^[1]; Yan, Jingwei ^[1]; Benites, Veronica T. ^[1]; Cheng-Yue, Clarabelle ^[1]; Tran, Thu ^[1];

^[1]; Mukhopadhyay, Aindrila ^[1];

^[2]; Scheller, Henrik V. ^[3]; Loqué, Dominique ^[4]

Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Division
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Division; Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering, Dept. of Chemical and Biomolecular Engineering
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Division; Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology
Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Division; Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology; National Inst. of Applied Sciences (INSA), Lyon (France)

Tight control and multifactorial regulation of gene expression are important challenges in genetic engineering and are critical for the development of regulatory circuits. In meeting these challenges we will facilitate transgene expression regulation and support the fine-tuning of metabolic pathways to avoid the accumulation of undesired intermediates. By employing the endoribonuclease Csy4 and its recognition sequence from Pseudomonas aeruginosa and manipulating 5'UTR of mRNA, we developed a two-component expression–repression system to tightly control synthesis of transgene products. We demonstrated that this regulatory device was functional in monocotyledonous and dicotyledonous plant species, and showed that it can be used to repress transgene expression by >400-fold and to synchronize transgene repression. In addition to tissue-specific transgene repression, this system offers stimuli-dependent expression control. Here, we identified 54 orthologous systems from various bacteria, using a bioinformatics approach and then validated in planta the activity for a few of those systems, demonstrating the potential diversity of such a two-component repressor system.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1361519

Journal Information:: ACS Synthetic Biology, Vol. 6, Issue 5; ISSN 2161-5063

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 10 works

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60 APPLIED LIFE SCIENCES
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gene regulation
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two-component repressor system

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