A Synthetic-Biology-Inspired Therapeutic Strategy for Targeting and Treating Hepatogenous Diabetes

Shao

et al. 2017

Quant Biol

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Background: Recently, optogenetics based on genetically encoded photosensitive proteins has emerged as an innovative technology platform to revolutionize manipulation of cellular behavior through light stimulation. It has enabled user defined control of various cellular behaviors with spatiotemporal precision and minimal invasiveness, creating unprecedented opportunities for biomedical applications. Results: This article reviews current advances in optogenetic networks designed for the treatment of human diseases. We highlight the advantages of these optogenetic networks, as well as emerging questions and future perspectives. Conclusions: Various optogenetic systems have been engineered to control biological processes at all levels using light and applied for numerous diseases, such as metabolic disorders, cancer, and immune diseases. Continued development of optogenetic modules will be necessary to precisely control of gene expression magnitude towards clinical medical practice in the context of real-world problems.

Section: Discussionmentioning

confidence: 99%

Engineering synthetic optogenetic networks for biomedical applications

Shao

et al. 2017

Quant Biol

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“…The expression of human placental SEAP in culture supernatants was determined using a p -nitrophenylphosphate-based light absorbance time course assay 51 . Briefly, 120 µL substrate solution [100 µL 2 × SEAP buffer containing 20 mM homoarginine, 1 mM MgCl 2 , 21% (vol/vol) diethanolamine, pH 9.8, and 20 µL substrate solution containing 120 mM p -nitrophenylphosphate] was added to 80 µL of heat-inactivated (65 °C, 30 min) cell culture supernatant.…”

Section: Methodsmentioning

confidence: 99%

A non-invasive far-red light-induced split-Cre recombinase system for controllable genome engineering in mice

Yang

et al. 2020

Nat Commun

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The Cre- loxP recombination system is a powerful tool for genetic manipulation. However, there are widely recognized limitations with chemically inducible Cre- loxP systems, and the UV and blue-light induced systems have phototoxicity and minimal capacity for deep tissue penetration. Here, we develop a far-red light-induced split Cre- loxP system (FISC system) based on a bacteriophytochrome optogenetic system and split-Cre recombinase, enabling optogenetical regulation of genome engineering in vivo solely by utilizing a far-red light (FRL). The FISC system exhibits low background and no detectable photocytotoxicity, while offering efficient FRL-induced DNA recombination. Our in vivo studies showcase the strong organ-penetration capacity of FISC system, markedly outperforming two blue-light-based Cre systems for recombination induction in the liver. Demonstrating its strong clinical relevance, we successfully deploy a FISC system using adeno-associated virus (AAV) delivery. Thus, the FISC system expands the optogenetic toolbox for DNA recombination to achieve spatiotemporally controlled, non-invasive genome engineering in living systems.

“…The production of human placental SEAP in cell-culture medium was assayed using a p-nitro phenyl phosphate-based light-absorbance time-course method as described previously ( 22 , 43 ). Briefly, 120 µL of substrate solution [100 µL of 2× SEAP assay buffer including 20 mM homoarginine, 1 mM MgCl 2 , 21% (wt/vol) diethanolamine, pH 9.8] and 20 µL of substrate solution containing 120 mM p-nitro phenyl phosphate was added to 80 µL of heat-deactivated (65 °C, 30 min) cell-culture supernatant, and the light absorbance was measured at 405 nm (37 °C) for 30 min using a Synergy H1 hybrid multimode microplate reader (BioTek Instruments, Inc.) using Gen5 software (version: 2.04).…”

Section: Methodsmentioning

confidence: 99%

Synthetic far-red light-mediated CRISPR-dCas9 device for inducing functional neuronal differentiation

Shao

Proc. Natl. Acad. Sci. U.S.A.

et al. 2018

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135

123

SignificanceWe have developed an optogenetic far-red light (FRL)-activated CRISPR-dCas9 system (FACE) that is orthogonal, fine-tunable, reversible, and has robust endogenous gene-activation profiles upon stimulation with FRL, with deep tissue penetration capacity, low brightness, short illumination time, and negligible phototoxicity. The FACE device is biocompatible and meets the criteria for safe medical application in humans, providing a robust differentiation strategy for mass production of functional neural cells from induced pluripotent stem cells simply by utilizing a beam of FRL. This optogenetic device has expanded the optogenetic toolkit for precise mammalian genome engineering in many areas of basic and translational research that require precise spatiotemporal control of cellular behavior, which may in turn boost the clinical progress of optogenetics-based precision therapy.