Switchable Cas9

Richter, Florian; Fonfara, Ines; Gelfert, Renate; Nack, Jennifer; Charpentier, Emmanuelle; Möglich, Andreas

doi:10.1016/j.copbio.2017.03.025

Cited by 38 publications

(32 citation statements)

References 55 publications

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“…Off-target effects can introduce unexpected mutations with serious consequences, and great efforts have been made to enhance the efficacy of CRISPR gene editing [96][97][98][99][100]. Many improvements, such as the use of chemical, temperature, and lightcontrolled stimuli, have demonstrated the ability to regulate the CRISPR-Cas system, but these modifications are not always ideal for every situation [101][102][103].…”

Section: Biotechnology Applications Of Acrs Against Type II and Type mentioning

confidence: 99%

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering

2019

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Clustered regularly interspaced short palindromic repeats (CRISPR)‐Cas adaptive immune defense systems, which are widely distributed in bacteria and Archaea, can provide sequence‐specific protection against foreign DNA or RNA in some cases. However, the evolution of defense systems in bacterial hosts did not lead to the elimination of phages, and some phages carry anti‐CRISPR genes that encode products that bind to the components mediating the defense mechanism and thus antagonize CRISPR‐Cas immune systems of bacteria. Given the extensive application of CRISPR‐Cas9 technologies in gene editing, in this review, we focus on the anti‐CRISPR proteins (Acrs) that inhibit CRISPR‐Cas systems for gene editing. We describe the discovery of Acrs in immune systems involving type I, II, and V CRISPR‐Cas immunity, discuss the potential function of Acrs in inactivating type II and V CRISPR‐Cas systems for gene editing and gene modulation, and provide an outlook on the development of important biotechnology tools for genetic engineering using Acrs.

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Section: Biotechnology Applications Of Acrs Against Type II and Type mentioning

confidence: 99%

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering

2019

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Section: Introductionmentioning

confidence: 99%

“…In addition to engineering CRISPR-Cas9 for diverse applications, there has also been broad interest in developing strategies to regulate CRISPR-Cas9 activity [8,9]. Such strategies promise to mitigate offtarget effects and allow the study of complex biological perturbations that require temporal or spatial resolution [9]. To date, most of the progress in this area has been focused on switching the activity of the Cas9 protein using chemical [10][11][12][13][14][15][16] or optical [17][18][19] inputs.…”

Section: Introductionmentioning

confidence: 99%

Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs

Kundert

Lucas

Watters

et al. 2018

Preprint

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The CRISPR-Cas9 system provides the ability to edit, repress, activate, or mark any gene (or DNA element) by pairing of a programmable single guide RNA (sgRNA) with a complementary sequence on the DNA target. Here we present a new method for small-molecule control of CRISPR-Cas9 function through insertion of RNA aptamers into the sgRNA. We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands. Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.

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“…In particular, CRISPR associated protein 9 (Cas9) is specifically appealing due to its high activity and flexibility in experimental design (Wang et al 2017;. Moreover, recent advances in protein engineering have yielded switchable Cas9 variants, whose activity can be controlled with spatiotemporal resolution by an external stimulus (Gangopadhyay et al 2019;Nihongaki et al 2018;Richter et al 2017;Zhou and Deiters 2016), and we envisage that such variants developed by us (Suzuki et al 2018) and others (Zetsche et al 2015;Davis et al 2015;Oakes et al 2016;Liu et al 2016;Nguyen et al 2016;Tang et al 2017;Senturk et al 2017;Rose et al 2017;Nihongaki et al 2015;Hemphill et al 2015;Richter et al 2016;Jain et al 2016;Zhou et al 2018) can benefit clinical development of in vivo gene editing.…”

mentioning

confidence: 99%

Applying switchable Cas9 variants to in vivo gene editing for therapeutic applications

et al. 2019

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Progress in targeted gene editing by programmable endonucleases has paved the way for their use in gene therapy. Particularly, Cas9 is an endonuclease with high activity and flexibility, rendering it an attractive option for therapeutic applications in clinical settings. Many disease-causing mutations could potentially be corrected by this versatile new technology. In addition, recently developed switchable Cas9 variants, whose activity can be controlled by an external stimulus, provide an extra level of spatiotemporal control on gene editing and are particularly desirable for certain applications. Here, we discuss the considerations and difficulties for implementing Cas9 to in vivo gene therapy. We put particular emphasis on how switchable Cas9 variants may resolve some of these barriers and advance gene therapy in the clinical setting.

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Switchable Cas9

Cited by 38 publications

References 55 publications

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering

Anti‐CRISPR proteins targeting the CRISPR‐Cas system enrich the toolkit for genetic engineering

Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs

Applying switchable Cas9 variants to in vivo gene editing for therapeutic applications

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