Engineered UV-A light-responsive gene expression system for measuring sun cream efficacy in mammalian cell culture

Wieland, Markus; Müller, Marius; Kyburz, Andreas; Heissig, Phillip; Wekenmann, Sebastian; Stolz, Franziska; Ausländer, Simon; Fussenegger, Martin

doi:10.1016/j.jbiotec.2014.09.008

Cited by 5 publications

(4 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Proteases can be adapted to operate in several environmental conditions and optimized to function in different organisms. Viral proteases such as tobacco etch virus protease (TEVp) are widely used in biotechnology for endoproteolytic removal of affinity tags from recombinant proteins 24 , and in synthetic biology to help convert extracellular or intracellular ligand sensing into transcriptional output 15 – 17 , 25 . They are also used to control protein levels via an N-terminal degron fused to the target protein with a protease-responsive cleavage site, as demonstrated in bacteria 14 .…”

Section: Introductionmentioning

confidence: 99%

Engineering protein-protein devices for multilayered regulation of mRNA translation using orthogonal proteases in mammalian cells

et al. 2018

View full text Add to dashboard Cite

The development of RNA-encoded regulatory circuits relying on RNA-binding proteins (RBPs) has enhanced the applicability and prospects of post-transcriptional synthetic network for reprogramming cellular functions. However, the construction of RNA-encoded multilayer networks is still limited by the availability of composable and orthogonal regulatory devices. Here, we report on control of mRNA translation with newly engineered RBPs regulated by viral proteases in mammalian cells. By combining post-transcriptional and post-translational control, we expand the operational landscape of RNA-encoded genetic circuits with a set of regulatory devices including: i) RBP-protease, ii) protease-RBP, iii) protease–protease, iv) protein sensor protease-RBP, and v) miRNA-protease/RBP interactions. The rational design of protease-regulated proteins provides a diverse toolbox for synthetic circuit regulation that enhances multi-input information processing-actuation of cellular responses. Our approach enables design of artificial circuits that can reprogram cellular function with potential benefits as research tools and for future in vivo therapeutics and biotechnological applications.

show abstract

Section: Introductionmentioning

confidence: 99%

Engineering protein-protein devices for multilayered regulation of mRNA translation using orthogonal proteases in mammalian cells

et al. 2018

View full text Add to dashboard Cite

show abstract

“…The sensing modules are based on intracellular antibodies (intrabodies) that have recently emerged as a new tool for therapeutic and functional genomics applications due to their ability to bind a wide range of proteins in several subcellular locations 11 . The actuation module takes advantage of the Tango-TEV technology, previously demonstrated to efficiently convert ligand/protein-induced dimerization into transcriptional output 12 – 14 (Supplementary Note 1, Supplementary Fig. 2 ).…”

Section: Resultsmentioning

confidence: 99%

Engineering modular intracellular protein sensor-actuator devices

et al. 2018

View full text Add to dashboard Cite

Understanding and reshaping cellular behaviors with synthetic gene networks requires the ability to sense and respond to changes in the intracellular environment. Intracellular proteins are involved in almost all cellular processes, and thus can provide important information about changes in cellular conditions such as infections, mutations, or disease states. Here we report the design of a modular platform for intrabody-based protein sensing-actuation devices with transcriptional output triggered by detection of intracellular proteins in mammalian cells. We demonstrate reporter activation response (fluorescence, apoptotic gene) to proteins involved in hepatitis C virus (HCV) infection, human immunodeficiency virus (HIV) infection, and Huntington’s disease, and show sensor-based interference with HIV-1 downregulation of HLA-I in infected T cells. Our method provides a means to link varying cellular conditions with robust control of cellular behavior for scientific and therapeutic applications.

show abstract

“…Upon UV light exposure, the cryptochrome CRY2 and the C1BN domain undergo dimerization. Wieland et al took advantage of this process, rewiring it to induce the production of a reporter 55 to detect the ability of various sunscreens to block harmful UV light. In the absence of light, both of the domains are expressed on the cell membrane.…”

Section: Cell-based Screening Systemsmentioning

confidence: 99%

Synthetic biology: applying biological circuits beyond novel therapies

Dobrin

Saxena

Fussenegger

2015

Integrative Biology

Self Cite

View full text Add to dashboard Cite

Synthetic biology, an engineering, circuit-driven approach to biology, has developed whole new classes of therapeutics. Unfortunately, these advances have thus far been undercapitalized upon by basic researchers. As discussed herein, using synthetic circuits, one can undertake exhaustive investigations of the endogenous circuitry found in nature, develop novel detectors and better temporally and spatially controlled inducers. One could detect changes in DNA, RNA, protein or even transient signaling events, in cell-based systems, in live mice, and in humans. Synthetic biology has also developed inducible systems that can be induced chemically, optically or using radio waves. This induction has been re-wired to lead to changes in gene expression, RNA stability and splicing, protein stability and splicing, and signaling via endogenous pathways. Beyond simple detectors and inducible systems, one can combine these modalities and develop novel signal integration circuits that can react to a very precise pre-programmed set of conditions or even to multiple sets of precise conditions. In this review, we highlight some tools that were developed in which these circuits were combined such that the detection of a particular event automatically triggered a specific output. Furthermore, using novel circuit-design strategies, circuits have been developed that can integrate multiple inputs together in Boolean logic gates composed of up to 6 inputs. We highlight the tools available and what has been developed thus far, and highlight how some clinical tools can be very useful in basic science. Most of the systems that are presented can be integrated together; and the possibilities far exceed the number of currently developed strategies.

show abstract

Engineered UV-A light-responsive gene expression system for measuring sun cream efficacy in mammalian cell culture

Cited by 5 publications

References 21 publications

Engineering protein-protein devices for multilayered regulation of mRNA translation using orthogonal proteases in mammalian cells

Engineering protein-protein devices for multilayered regulation of mRNA translation using orthogonal proteases in mammalian cells

Engineering modular intracellular protein sensor-actuator devices

Synthetic biology: applying biological circuits beyond novel therapies

Contact Info

Product

Resources

About