Rui Miao
@kth.se
Researcher at Department of CBH, division of System biology
Royal Institute of Technology
RESEARCH, TEACHING, or OTHER INTERESTS
Biotechnology, Biochemistry, Genetics and Molecular Biology, Plant Science, Agricultural and Biological Sciences
Scopus Publications
Scopus Publications
Rui Miao, Michael Jahn, Kiyan Shabestary, Gilles Peltier, and Elton P Hudson
Oxford University Press (OUP)
Abstract Barcoded mutant libraries are a powerful tool for elucidating gene function in microbes, particularly when screened in multiple growth conditions. Here, we screened a pooled CRISPR interference library of the model cyanobacterium Synechocystis sp. PCC 6803 in 11 bioreactor-controlled conditions, spanning multiple light regimes and carbon sources. This gene repression library contained 21,705 individual mutants with high redundancy over all open reading frames and noncoding RNAs. Comparison of the derived gene fitness scores revealed multiple instances of gene repression being beneficial in 1 condition while generally detrimental in others, particularly for genes within light harvesting and conversion, such as antennae components at high light and PSII subunits during photoheterotrophy. Suboptimal regulation of such genes likely represents a tradeoff of reduced growth speed for enhanced robustness to perturbation. The extensive data set assigns condition-specific importance to many previously unannotated genes and suggests additional functions for central metabolic enzymes. Phosphoribulokinase, glyceraldehyde-3-phosphate dehydrogenase, and the small protein CP12 were critical for mixotrophy and photoheterotrophy, which implicates the ternary complex as important for redirecting metabolic flux in these conditions in addition to inactivation of the Calvin cycle in the dark. To predict the potency of sgRNA sequences, we applied machine learning on sgRNA sequences and gene repression data, which showed the importance of C enrichment and T depletion proximal to the PAM site. Fitness data for all genes in all conditions are compiled in an interactive web application.
Ian S. Yunus, Josefine Anfelt, Emil Sporre, Rui Miao, Elton P. Hudson, and Patrik R. Jones
Elsevier BV
Kiyan Shabestary, Hugo Pineda Hernández, Rui Miao, Emil Ljungqvist, Olivia Hallman, Emil Sporre, Filipe Branco dos Santos, and Elton P. Hudson
Elsevier BV
Xufeng Liu, Hao Xie, Stamatina Roussou, Rui Miao, and Peter Lindblad
De Gruyter
Rui Miao, Hao Xie, Xufeng Liu, Pia Lindberg, and Peter Lindblad
Elsevier BV
Xufeng Liu, Rui Miao, Pia Lindberg, and Peter Lindblad
Royal Society of Chemistry (RSC)
Systematic modular engineering of a photosynthetic microorganism resulted in cells with significant levels and rates of 1-butanol production directly from CO2.
Kamonchanock Eungrasamee, Rui Miao, Aran Incharoensakdi, Peter Lindblad, and Saowarath Jantaro
Springer Science and Business Media LLC
Rui Miao, Hao Xie, and Peter Lindblad
Springer Science and Business Media LLC
Rui Miao, Hao Xie, Felix M. Ho, and Peter Lindblad
Elsevier BV
Rui Miao, Xufeng Liu, Elias Englund, Pia Lindberg, and Peter Lindblad
Elsevier BV
Rui Miao, Adam Wegelius, Claudia Durall, Feiyan Liang, Namita Khanna, and Peter Lindblad
Springer International Publishing
Elias Englund, Johan Andersen-Ranberg, Rui Miao, Björn Hamberger, and Pia Lindberg
American Chemical Society (ACS)
Forskolin is a high value diterpenoid with a broad range of pharmaceutical applications, naturally found in root bark of the plant Coleus forskohlii. Because of its complex molecular structure, chemical synthesis of forskolin is not commercially attractive. Hence, the labor and resource intensive extraction and purification from C. forskohlii plants remains the current source of the compound. We have engineered the unicellular cyanobacterium Synechocystis sp. PCC 6803 to produce the forskolin precursor 13R-manoyl oxide (13R-MO), paving the way for light driven biotechnological production of this high value compound. In the course of this work, a new series of integrative vectors for use in Synechocystis was developed and used to create stable lines expressing chromosomally integrated CfTPS2 and CfTPS3, the enzymes responsible for the formation of 13R-MO in C. forskohlii. The engineered strains yielded production titers of up to 0.24 mg g–1 DCW 13R-MO. To increase the yield, 13R-MO producing strains were further engineered by introduction of selected enzymes from C. forskohlii, improving the titer to 0.45 mg g–1 DCW. This work forms a basis for further development of production of complex plant diterpenoids in cyanobacteria.