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A single promoter-TALE system for tissue-specific and tuneable expression of multiple genes in rice.

Published version
Peer-reviewed

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Article

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Authors

Hua, Lei 
Vlad, Daniela 

Abstract

In biological discovery and engineering research, there is a need to spatially and/or temporally regulate transgene expression. However, the limited availability of promoter sequences that are uniquely active in specific tissue-types and/or at specific times often precludes co-expression of multiple transgenes in precisely controlled developmental contexts. Here, we developed a system for use in rice that comprises synthetic designer transcription activator-like effectors (dTALEs) and cognate synthetic TALE-activated promoters (STAPs). The system allows multiple transgenes to be expressed from different STAPs, with the spatial and temporal context determined by a single promoter that drives expression of the dTALE. We show that two different systems-dTALE1-STAP1 and dTALE2-STAP2-can activate STAP-driven reporter gene expression in stable transgenic rice lines, with transgene transcript levels dependent on both dTALE and STAP sequence identities. The relative strength of individual STAP sequences is consistent between dTALE1 and dTALE2 systems but differs between cell-types, requiring empirical evaluation in each case. dTALE expression leads to off-target activation of endogenous genes but the number of genes affected is substantially less than the number impacted by the somaclonal variation that occurs during the regeneration of transformed plants. With the potential to design fully orthogonal dTALEs for any genome of interest, the dTALE-STAP system thus provides a powerful approach to fine-tune the expression of multiple transgenes, and to simultaneously introduce different synthetic circuits into distinct developmental contexts.

Description

Funder: University of Oxford


Funder: Bill & Melinda Gates Foundation; Id: http://dx.doi.org/10.13039/100000865


Funder: Academia Sinica; Id: http://dx.doi.org/10.13039/501100001869


Funder: Ministry of Education; Id: http://dx.doi.org/10.13039/501100002701


Funder: Royal Society; Id: http://dx.doi.org/10.13039/501100000288


Funder: BBSRC; Id: http://dx.doi.org/10.13039/501100000268


Funder: Australian National University; Id: http://dx.doi.org/10.13039/501100000995

Keywords

cell-type-specific gene expression, dTALE-STAP, rice, synthetic gene circuits, Genes, Reporter, Oryza, Plants, Plants, Genetically Modified, Promoter Regions, Genetic, Transgenes

Journal Title

Plant Biotechnol J

Conference Name

Journal ISSN

1467-7644
1467-7652

Volume Title

Publisher

Wiley
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/P003117/1)