Open AccessEndoribonuclease-Based Two-Component Repressor Systems for Tight Gene Expression Control in Plants
Author(s) -
Yan Liang,
Sarah M. Richardson,
Jingwei Yan,
Veronica T. Benites,
Clarabelle Cheng-Yue,
Thu Hương Trần,
Jenny C. Mortimer,
Aindrila Mukhopadhyay,
Jay D. Keasling,
Henrik Vibe Scheller,
Dominique Loqué
Publication year - 2017
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.6b00295
Subject(s) - repressor , endoribonuclease , component (thermodynamics) , gene , biology , genetics , gene expression , computational biology , rna , physics , rnase p , thermodynamics
Tight control and multifactorial regulation of gene expression are important challenges in genetic engineering and are critical for the development of regulatory circuits. Meeting these challenges 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. Using a bioinformatics approach, we identified 54 orthologous systems from various bacteria, and then validated in planta the activity for a few of those systems, demonstrating the potential diversity of such a two-component repressor system.