Expanding the genetic code of Drosophila melanogaster

Bianco, Ambra; Townsley, Fiona M.; Greiss, Sebastian; Lang, Kathrin; Chin, Jason W.

doi:10.1038/nchembio.1043

Cited by 190 publications

(159 citation statements)

References 24 publications

Supporting

Mentioning

155

Contrasting

Unclassified

Order By: Relevance

“…ND, no data. mammalian culture cells, and plants (58)(59)(60)(61)(62). Similar translational switches can also be constructed using these pairs and can be easily implemented with applicable organisms.…”

Section: Discussionmentioning

confidence: 99%

High-Yield, Zero-Leakage Expression System with a Translational Switch Using Site-Specific Unnatural Amino Acid Incorporation

Minaba

Kato

2014

Appl Environ Microbiol

View full text Add to dashboard Cite

Synthetic biologists construct complex biological circuits by combinations of various genetic parts. Many genetic parts that are orthogonal to one another and are independent of existing cellular processes would be ideal for use in synthetic biology. However, our toolbox is still limited with respect to the bacterium Escherichia coli, which is important for both research and industrial use. The site-specific incorporation of unnatural amino acids is a technique that incorporates unnatural amino acids into proteins using a modified exogenous aminoacyl-tRNA synthetase/tRNA pair that is orthogonal to any native pairs in a host and is independent from other cellular functions. Focusing on the orthogonality and independency that are suitable for the genetic parts, we designed novel AND gate and translational switches using the unnatural amino acid 3-iodo-L-tyrosine incorporation system in E. coli. A translational switch was turned on after addition of 3-iodo-L-tyrosine in the culture medium within minutes and allowed tuning of switchability and translational efficiency. As an application, we also constructed a gene expression system that produced large amounts of proteins under induction conditions and exhibited zero-leakage expression under repression conditions. Similar translational switches are expected to be applicable also for eukaryotes such as yeasts, nematodes, insects, mammalian cells, and plants.

show abstract

Section: Discussionmentioning

confidence: 99%

High-Yield, Zero-Leakage Expression System with a Translational Switch Using Site-Specific Unnatural Amino Acid Incorporation

Minaba

Kato

2014

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Later, Carell and co-workers showed that a norbornenemodified Lys (68) can be genetically encoded into His 6 -tagged human polymerase κ, and that efficient labeling of polymerase κ can be accomplished with the nitrile imines generated from either the hydrazonyl chloride precursor or the corresponding tetrazole [23]. The utility of the norbornene modified amino acids was also demonstrated in the tetrazine ligation in eukaryotic cells by other research groups [46][47][48]. Smaller strained alkenes such as cyclopropene have also been developed for the photoclick chemistry.…”

Section: Site-specific Labeling Of Proteins Via the Photoinduced Tetrmentioning

confidence: 94%

Photo-Triggered Click Chemistry for Biological Applications

Herner¹,

Qi²

2015

Cycloadditions in Bioorthogonal Chemistry

View full text Add to dashboard Cite

In the last decade and a half, numerous bioorthogonal reactions have been developed with a goal to study biological processes in their native environment, i.e., in living cells and animals. Among them, the photo-triggered reactions offer several unique advantages including operational simplicity with the use of light rather than toxic metal catalysts and ligands, and exceptional spatiotemporal control through the application of an appropriate light source with pre-selected wavelength, light intensity and exposure time. While the photoinduced reactions have been studied extensively in materials research, e.g., on macromolecular surface, the adaptation of these reactions for chemical biology applications is still in its infancy. In this chapter, we review the recent efforts in the discovery and optimization the photo-triggered bioorthogonal reactions, with a focus on those that have shown broad utility in biological systems. We discuss in each cases the chemical and mechanistic background, the kinetics of the reactions and the biological applicability together with the limiting factors.

show abstract

“…This technology made it possible to photocontrol protein localization 45 , signal transduction 46,47 and gene expression 49 . This approach, demonstrated initially in mammalian cells, should rapidly benefit from the recent upgrade of the unnatural mutagenesis strategy to multicellular organisms 50 .…”

Section: Chemical and Genetic Hybrid Photoactuatorsmentioning

confidence: 99%

How to control proteins with light in living systems

et al. 2014

View full text Add to dashboard Cite

The possibility offered by photocontrolling the activity of biomolecules in vivo while recording physiological parameters is opening up new opportunities for the study of physiological processes at the single-cell level in a living organism. For the last decade, such tools have been mainly used in neuroscience, and their application in freely moving animals has revolutionized this field. New photochemical approaches enable the control of various cellular processes by manipulating a wide range of protein functions in a noninvasive way and with unprecedented spatiotemporal resolution. We are at a pivotal moment where biologists can adapt these cutting-edge technologies to their system of study. This user-oriented review presents the state of the art and highlights technical issues to be resolved in the near future for wide and easy use of these powerful approaches.

show abstract

Expanding the genetic code of Drosophila melanogaster

Cited by 190 publications

References 24 publications

High-Yield, Zero-Leakage Expression System with a Translational Switch Using Site-Specific Unnatural Amino Acid Incorporation

High-Yield, Zero-Leakage Expression System with a Translational Switch Using Site-Specific Unnatural Amino Acid Incorporation

Photo-Triggered Click Chemistry for Biological Applications

How to control proteins with light in living systems

Contact Info

Product

Resources

About