CRISPR–Cas9-based photoactivatable transcription systems to induce neuronal differentiation

Nihongaki, Yuta; Furuhata, Yuichi; Otabe, Takahiro; Hasegawa, Shunsuke; Yoshimoto, Keitaro; Sato, Moritoshi

doi:10.1038/nmeth.4430

Cited by 152 publications

(148 citation statements)

References 20 publications

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“…Investigating the functions of essential genes is possible through the use of inducible CRISPRi systems, eliminating the need to construct thermo‐sensitive or other conditional mutants . Another advantage of CRISPR‐Cas systems for gene expression manipulation is the ability to precisely control and fine‐tune expression through use of titratable promoters and photoactivatable systems . The inducible and titratable nature of these systems allows to design genetic circuits that respond to environmental stimuli for engineering of industrial strains, which can undergo a metabolic shift from production to growth phase or control accumulation of metabolites .…”

Section: Resultsmentioning

confidence: 99%

CRISPR‐Enabled Tools for Engineering Microbial Genomes and Phenotypes

Tarasava

Eckert

et al. 2018

Biotechnology Journal

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In recent years CRISPR-Cas technologies have revolutionized microbial engineering approaches. Genome editing and non-editing applications of various CRISPR-Cas systems have expanded the throughput and scale of engineering efforts, as well as opened up new avenues for manipulating genomes of non-model organisms. As we expand the range of organisms used for biotechnological applications, we need to develop better, more versatile tools for manipulation of these systems. Here the authors summarize the current advances in microbial gene editing using CRISPR-Cas based tools and highlight state-of-the-art methods for high-throughput, efficient genome-scale engineering in model organisms Escherichia coli and Saccharomyces cerevisiae. The authors also review non-editing CRISPR-Cas applications available for gene expression manipulation, epigenetic remodeling, RNA editing, labeling, and synthetic gene circuit design. Finally, the authors point out the areas of research that need further development in order to expand the range of applications and increase the utility of these new methods.

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Section: Resultsmentioning

confidence: 99%

CRISPR‐Enabled Tools for Engineering Microbial Genomes and Phenotypes

Tarasava

Eckert

et al. 2018

Biotechnology Journal

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“…60 Recently, CRISPR-Cas9-based photoactivatable transcription system (CPTS2.0) using padCas9-SAM was able to robustly activate gene transcription level in a light-dependent manner in human cell lines. 72…”

Section: Guide Rna Modification For Spatiotemporal Regulationmentioning

confidence: 99%

CRISPR‐Cas system: Toward a more efficient technology for genome editing and beyond

Ahmadzadeh

Farajnia

Baghban

et al. 2019

J of Cellular Biochemistry

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Genome engineering technology is of great interest for biomedical research that enables scientists to make specific manipulation in the DNA sequence. Early methods for introducing double‐stranded DNA breaks relies on protein‐based systems. These platforms have enabled fascinating advances, but all are costly and time‐consuming to engineer, preventing these from gaining high‐throughput applications. The CRISPR‐Cas9 system, co‐opted from bacteria, has generated considerable excitement in gene targeting. In this review, we describe gene targeting techniques with an emphasis on recent strategies to improve the specificities of CRISPR‐Cas systems for nuclease and non‐nuclease applications.

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“…Fusion of pMag to an inter‐SH2 domain of phosphatidylinositol 3‐kinase, for instance, enabled light‐dependent recruitment to a membrane‐tethered nMag variant, thereby activating PI(3,4,5)P 3 signaling. Also, fusion of the magnets to the N‐ and C‐terminal fragments of a split Streptococcus pyogenes CRISPR/Cas9 or split T7 polymerase allowed for tight light control of genome editing and gene expression, respectively . A notable property of the Magnets system is that dimerization requires excitation of both binding partners—nMag and pMag.…”

Section: Gaining Optogenetic Control With Lov Domainsmentioning

confidence: 99%

Controlling Cells with Light and LOV

et al. 2018

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Optogenetics is a powerful method for studying dynamic processes in living cells and has advanced cell biology research over the recent past. Key to the successful application of optogenetics is the careful design of the light‐sensing module, typically employing a natural or engineered photoreceptor that links the exogenous light input to the cellular process under investigation. Light–oxygen–voltage (LOV) domains, a highly diverse class of small blue light sensors, have proven to be particularly versatile for engineering optogenetic input modules. These can function via diverse modalities, including inducible allostery, protein recruitment, dimerization, or dissociation. This study reviews recent advances in the development of LOV domain‐based optogenetic tools and their application for studying and controlling selected cellular functions. Focusing on the widely employed LOV2 domain from Avena sativa phototropin‐1, this review highlights the broad spectrum of engineering opportunities that can be explored to achieve customized optogenetic regulation. Finally, major bottlenecks in the development of optogenetic methods are discussed and strategies to overcome these with recent synthetic biology approaches are pointed out.

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CRISPR–Cas9-based photoactivatable transcription systems to induce neuronal differentiation

Cited by 152 publications

References 20 publications

CRISPR‐Enabled Tools for Engineering Microbial Genomes and Phenotypes

CRISPR‐Enabled Tools for Engineering Microbial Genomes and Phenotypes

CRISPR‐Cas system: Toward a more efficient technology for genome editing and beyond

Controlling Cells with Light and LOV

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