@article{67603a058c224fdab5ec6b924162c3f7,
title = "A single promoter-TALE system for tissue-specific and tuneable expression of multiple genes in rice",
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.",
keywords = "cell-type-specific gene expression, dTALE-STAP, rice, synthetic gene circuits",
author = "Florence Danila and Tom Schreiber and Maria Ermakova and Lei Hua and Daniela Vlad and Lo, {Shuen Fang} and Chen, {Yi Shih} and Julia Lambret-Frotte and Hermanns, {Anna S.} and Benedikt Athmer and {von Caemmerer}, Susanne and Yu, {Su May} and Hibberd, {Julian M.} and Alain Tissier and Furbank, {Robert T.} and Steven Kelly and Langdale, {Jane A.}",
note = "Funding Information: The authors thank Xueqin Wang, Riya Kuruvilla (ANU), and Na Wang (Cambridge) for technical assistance and Robert Sharwood for the gifts of antibodies. The authors thank the Australian Plant Phenomics Facility supported under the National Collaborative Research Infrastructure Strategy of the Australian Government and also acknowledge the instruments and expertise of Microscopy Australia at the Centre for Advanced Microscopy, Australian National University, enabled by NCRIS, university, and state government support. TS, BA, and AT were funded in part by a BMBF grant (# 031B0548) from the German Federal Ministry for Education and Research to AT, and by core institute funding at IPB, Halle; JLF was funded by BBSRC sLOLA grant BB/P003117/1; SK is a Royal Society University Research Fellow; SFL was supported in part by the Advanced Plant Biotechnology Center from the Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan; YSC was supported by Academia Sinica; all other authors were funded by grants from the Bill & Melinda Gates Foundation to the University of Oxford—OPP1129902 (C Rice Phase III) and INV‐002970 (C Rice Phase IV). 4 4 Funding Information: The authors thank Xueqin Wang, Riya Kuruvilla (ANU), and Na Wang (Cambridge) for technical assistance and Robert Sharwood for the gifts of antibodies. The authors thank the Australian Plant Phenomics Facility supported under the National Collaborative Research Infrastructure Strategy of the Australian Government and also acknowledge the instruments and expertise of Microscopy Australia at the Centre for Advanced Microscopy, Australian National University, enabled by NCRIS, university, and state government support. TS, BA, and AT were funded in part by a BMBF grant (# 031B0548) from the German Federal Ministry for Education and Research to AT, and by core institute funding at IPB, Halle; JLF was funded by BBSRC sLOLA grant BB/P003117/1; SK is a Royal Society University Research Fellow; SFL was supported in part by the Advanced Plant Biotechnology Center from the Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan; YSC was supported by Academia Sinica; all other authors were funded by grants from the Bill & Melinda Gates Foundation to the University of Oxford—OPP1129902 (C4 Rice Phase III) and INV-002970 (C4 Rice Phase IV). Publisher Copyright: {\textcopyright} 2022 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.",
year = "2022",
month = sep,
doi = "10.1111/pbi.13864",
language = "English",
volume = "20",
pages = "1786--1806",
journal = "Plant Biotechnology Journal",
issn = "1467-7644",
publisher = "Wiley-Blackwell",
number = "9",
}