scholarly journals CreLite: An Optogenetically Controlled Cre/loxP System Using Red Light

2019 ◽  
Author(s):  
Shuo-Ting Yen ◽  
Kenneth A. Trimmer ◽  
Nader Aboul ◽  
Rachel D. Mullen ◽  
James C. Culver ◽  
...  
Keyword(s):  
Gene Expression ◽  
Light Exposure ◽  
Red Light ◽  
Regulatory Elements ◽  
Cell Labeling ◽  
P System ◽  
Zebrafish Embryos ◽  
Control Of Gene Expression ◽  
Spatial Control ◽  
Temporal And Spatial

ABSTRACTPrecise manipulation of gene expression with temporal and spatial control is essential for functional studies and the determination of cell lineage relationships in complex biological systems. The Cre-loxP system is commonly used for gene manipulation at desired times and places. However, specificity is dependent on the availability of tissue- or cell-specific regulatory elements used in combination with Cre or CreER (tamoxifen-inducible). Here we present CreLite, an optogenetically-controlled Cre system using red light in developing zebrafish embryos. Cre activity is disabled by splitting Cre and fusing the inactive halves with the Arabidopsis thaliana red light-inducible binding partners, PhyB and PIF6. In addition, PhyB-PIF6 binding requires phycocyanobilin (PCB), providing an additional layer of control. Upon exposure to red light (660 nm) illumination, the PhyB-CreC and PIF6-CreN fusion proteins come together in the presence of PCB to restore Cre activity. Red-light exposure of transgenic zebrafish embryos harboring a Cre-dependent multi-color fluorescent protein reporter (ubi:zebrabow) injected with CreLite mRNAs and PCB, resulted in Cre activity as measured by the generation of multi-spectral cell labeling in various tissues, including heart, skeletal muscle and epithelium. We show that CreLite can be used for gene manipulations in whole embryos or small groups of cells at different stages of development. CreLite provides a novel optogenetic tool for precise temporal and spatial control of gene expression in zebrafish embryos that may also be useful in cell culture, ex vivo organ culture, and other animal models for developmental biology studies.

scholarly journals Temporal and spatial control of gene expression in horticultural crops

2014 ◽  
Vol 1 (1) ◽  
Author(s):  
Manjul Dutt ◽  
Sadanand A Dhekney ◽  
Leonardo Soriano ◽  
Raju Kandel ◽  
Jude W Grosser
Keyword(s):  
Gene Expression ◽  
Control Of Gene Expression ◽  
Spatial Control ◽  
Horticultural Crops ◽  
Temporal And Spatial

scholarly journals Artificial complementary chromatic acclimation gene expression system in Escherichia coli

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Dwi Ariyanti ◽  
Kazunori Ikebukuro ◽  
Koji Sode
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Light Exposure ◽  
Expression System ◽  
Red Light ◽  
Green Light ◽  
Light Illumination ◽  
Multiple Gene ◽  
Gene Expression System ◽  
Chromatic Acclimation

Abstract Background The development of multiple gene expression systems, especially those based on the physical signals, such as multiple color light irradiations, is challenging. Complementary chromatic acclimation (CCA), a photoreversible process that facilitates the control of cellular expression using light of different wavelengths in cyanobacteria, is one example. In this study, an artificial CCA systems, inspired by type III CCA light-regulated gene expression, was designed by employing a single photosensor system, the CcaS/CcaR green light gene expression system derived from Synechocystis sp. PCC6803, combined with G-box (the regulator recognized by activated CcaR), the cognate cpcG2 promoter, and the constitutively transcribed promoter, the PtrcΔLacO promoter. Results One G-box was inserted upstream of the cpcG2 promoter and a reporter gene, the rfp gene (green light-induced gene expression), and the other G-box was inserted between the PtrcΔLacO promoter and a reporter gene, the bfp gene (red light-induced gene expression). The Escherichia coli transformants with plasmid-encoded genes were evaluated at the transcriptional and translational levels under red or green light illumination. Under green light illumination, the transcription and translation of the rfp gene were observed, whereas the expression of the bfp gene was repressed. Under red light illumination, the transcription and translation of the bfp gene were observed, whereas the expression of the rfp gene was repressed. During the red and green light exposure cycles at every 6 h, BFP expression increased under red light exposure while RFP expression was repressed, and RFP expression increased under green light exposure while BFP expression was repressed. Conclusion An artificial CCA system was developed to realize a multiple gene expression system, which was regulated by two colors, red and green lights, using a single photosensor system, the CcaS/CcaR system derived from Synechocystis sp. PCC6803, in E. coli. The artificial CCA system functioned repeatedly during red and green light exposure cycles. These results demonstrate the potential application of this CCA gene expression system for the production of multiple metabolites in a variety of microorganisms, such as cyanobacteria.

Control of gene expression using a red- and far-red light–responsive bi-stable toggle switch

2014 ◽  
Vol 9 (3) ◽  
pp. 622-632 ◽  
Author(s):  
Konrad Müller ◽  
Matias D Zurbriggen ◽  
Wilfried Weber
Keyword(s):  
Gene Expression ◽  
Red Light ◽  
Toggle Switch ◽  
Control Of Gene Expression

Vitamin D Control of Gene Expression: Temporal and Spatial Parameters for Organization of the Regulatory Machinery

2008 ◽  
Vol 18 (2) ◽  
pp. 163-172 ◽  
Author(s):  
Martin Montecino ◽  
Gary Stein ◽  
Janet L. Stein ◽  
Jane B. Lian ◽  
Andre J. van Wijnen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Vitamin D ◽  
Control Of Gene Expression ◽  
Regulatory Machinery ◽  
Spatial Parameters ◽  
Temporal And Spatial

scholarly journals Combining Transcriptomics and Proteomics Reveals Potential Post-transcriptional Control of Gene Expression After Light Exposure in Metarhizium acridum

2019 ◽  
Vol 9 (9) ◽  
pp. 2951-2961 ◽  
Author(s):  
Guilherme T. P. Brancini ◽  
Márcia E. S. Ferreira ◽  
Drauzio E. N. Rangel ◽  
Gilberto Ú. L. Braga
Keyword(s):  
Gene Expression ◽  
Transcriptional Control ◽  
Light Exposure ◽  
Control Of Gene Expression ◽  
Metarhizium Acridum

Phytochrome signal-transduction: characterization of pathways and isolation of mutants

1995 ◽  
Vol 350 (1331) ◽  
pp. 67-74 ◽  
Keyword(s):  
Gene Expression ◽  
Signal Transduction ◽  
Red Light ◽  
Signal Transduction Pathway ◽  
Regulatory Elements ◽  
Signal Transduction Pathways ◽  
Expression Of Genes ◽  
Genes Encoding ◽  
Regulated Gene Expression ◽  
Dependent Pathway

The study of phytochrome signalling has yielded a wealth of data describing both the perception of light by the receptor, and the terminal steps in phytochrome-regulated gene expression by a number of transcription factors. We are now focusing on establishing the intervening steps linking phytochrome photoactivation to gene expression, and the regulation and interactions of these signalling pathways. Recent work has utilized both a pharmacological approach in phototrophic soybean suspension cultures and microinjection techniques in tomato to establish three distinct phytochrome signal-transduction pathways: (i) a calcium-dependent pathway that regulates the expression of genes encoding the chlorophyll a/b binding protein ( CAB ) and other components of photosystem II; (ii) a cGMP-dependent pathway that regulates the expression of the gene encoding chalcone synthase ( CHS ) and the production of anthocyanin pigments; and (iii) a pathway dependent upon both calcium and cGMP that regulates the expression of genes encoding components of photosystem I and is necessary for the production of mature chloroplasts. To study the components and the regulation of phytochrome signal-transduction pathways, mutants with altered photomorphogenic responses have been isolated by a number of laboratories. However, with several possible exceptions, little real progress has been made towards the isolation of mutants in positive regulatory elements of the phytochrome signal-transduction pathway. We have characterized a novel phytochrome A (phyA)-mediated far-red light (FR) response in Arabidopsis seedlings which we are currently using to screen for specific phyA signal-transduction mutants.

scholarly journals Synthetic RNA switches as a tool for temporal and spatial control over gene expression

2012 ◽  
Vol 23 (5) ◽  
pp. 679-688 ◽  
Author(s):  
Andrew L Chang ◽  
Joshua J Wolf ◽  
Christina D Smolke
Keyword(s):  
Gene Expression ◽  
Spatial Control ◽  
Rna Switches ◽  
Temporal And Spatial
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