Epigenome Editing: State of the Art, Concepts, and Perspectives

Kungulovski, Goran; Jeltsch, Albert

doi:10.1016/j.tig.2015.12.001

Cited by 171 publications

(99 citation statements)

References 91 publications

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“…Increased specificity has been achieved using paired TALEN or CRISPR/Cas9 nickases , truncated gRNAs (Fu et al, 2014), or dimeric CRISPR/Cas9 nucleases (Guilinger et al, 2014;Tsai et al, 2014). Other diverse applications have emerged, including targeted regulation of gene expression, targeted epigenetic modification, or site-specific base editing through deamination (La Russa and Qi, 2015;Kungulovski and Jeltsch, 2016;Zong et al, 2017;Shimatani et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

et al. 2017

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We report a comprehensive toolkit that enables targeted, specific modification of monocot and dicot genomes using a variety of genome engineering approaches. Our reagents, based on transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, are systematized for fast, modular cloning and accommodate diverse regulatory sequences to drive reagent expression. Vectors are optimized to create either single or multiple gene knockouts and large chromosomal deletions. Moreover, integration of geminivirus-based vectors enables precise gene editing through homologous recombination. Regulation of transcription is also possible. A Web-based tool streamlines vector selection and construction. One advantage of our platform is the use of the Csy-type (CRISPR system yersinia) ribonuclease 4 (Csy4) and tRNA processing enzymes to simultaneously express multiple guide RNAs (gRNAs). For example, we demonstrate targeted deletions in up to six genes by expressing 12 gRNAs from a single transcript. Csy4 and tRNA expression systems are almost twice as effective in inducing mutations as gRNAs expressed from individual RNA polymerase III promoters. Mutagenesis can be further enhanced 2.5-fold by incorporating the Trex2 exonuclease. Finally, we demonstrate that Cas9 nickases induce gene targeting at frequencies comparable to native Cas9 when they are delivered on geminivirus replicons. The reagents have been successfully validated in tomato (Solanum lycopersicum), tobacco (Nicotiana tabacum), Medicago truncatula, wheat (Triticum aestivum), and barley (Hordeum vulgare).

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Section: Introductionmentioning

confidence: 99%

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

et al. 2017

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“…Los EpiEfectores son sustancias que comprenden segmentos de un polipéptido llamados dominios de reconocimiento de ADN, diseñados con dedo de zinc, TAL, o complejo CRISPR/Cas9 modificado (clustered regularly interspaced short palindromic repeats repeticiones palindrómicas cortas agrupadas y regularmente interespaciadas), es decir, modificaciones recientes de este sistema, que permiten actuar sobre la transcripción de los genes; además de los dominios catalíticos de una enzima modificadora de la cromatina son los necesarios para dirigir eficazmente los mecanismos epigenéticos (12).…”

Section: El Futurounclassified

“…Esto pone de manifiesto la importancia de las modificaciones de las histonas y la metilación del ADN en el dolor (12).…”

Section: El Futurounclassified

La epigenética en el tratamiento del dolor

Carrillo

Toro

Bolívar

et al. 2017

Rev. Soc. Esp. Dolor.

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epigenética en el tratamiento del dolor / Epigenetics in the treatment of pain. Rev. Soc. Esp. Dolor. 2017. doi: 10.20986/resed.2017.3620/2017 Este es un archivo PDF de un manuscrito inédito que ha sido aceptado para su publicación en la Revista de la Sociedad Española del Dolor. Como un servicio a nuestros clientes estamos proporcionando esta primera versión del manuscrito en estado de prepublicación. El manuscrito será sometido a la corrección de estilo final, composición y revisión de la prueba resultante antes de que se publique en su forma final. Tenga en cuenta que durante el proceso de producción se pueden dar errores lo que podría afectar el contenido final. El copyright y todos los derechos legales que se aplican al artículo pertenecen a la Revista de la Sociedad Española de Dolor. LA EPIGENÉTICA EN EL TRATAMIENTO DEL

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“…One of its most promising approaches is to overwrite epigenetic marks at specific genome regions by targeting epigenetic effector domains (EDs) fused to a DNA binding domain (DBD) that specifically recognizes a particular DNA sequence. 14 These chimeric proteins can be very useful in basic research 15 and might also have important applications in the study of diseases that show epigenetic alterations, such as cancer. 16,17 Recent studies have shown targeted DNA demethylation and gene reactivation in mammalian cells by fusing different enzymes involved in DNA demethylation to diverse DBDs.…”

Section: Introductionmentioning

confidence: 99%

Targeted DNA demethylation in human cells by fusion of a plant 5-methylcytosine DNA glycosylase to a sequence-specific DNA binding domain

2017

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DNA methylation is a crucial epigenetic mark associated to gene silencing, and its targeted removal is a major goal of epigenetic editing. In animal cells, DNA demethylation involves iterative 5mC oxidation by TET enzymes followed by replication-dependent dilution and/or replication-independent DNA repair of its oxidized derivatives. In contrast, plants use specific DNA glycosylases that directly excise 5mC and initiate its substitution for unmethylated C in a base excision repair process. In this work, we have fused the catalytic domain of Arabidopsis ROS1 5mC DNA glycosylase (ROS1_CD) to the DNA binding domain of yeast GAL4 (GBD). We show that the resultant GBD-ROS1_CD fusion protein binds specifically a GBDtargeted DNA sequence in vitro. We also found that transient in vivo expression of GBD-ROS1_CD in human cells specifically reactivates transcription of a methylation-silenced reporter gene, and that such reactivation requires both ROS1_CD catalytic activity and GBD binding capacity. Finally, we show that reactivation induced by GBD-ROS1_CD is accompanied by decreased methylation levels at several CpG sites of the targeted promoter. All together, these results show that plant 5mC DNA glycosylases can be used for targeted active DNA demethylation in human cells.

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Epigenome Editing: State of the Art, Concepts, and Perspectives

Cited by 171 publications

References 91 publications

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

A Multipurpose Toolkit to Enable Advanced Genome Engineering in Plants

La epigenética en el tratamiento del dolor

Targeted DNA demethylation in human cells by fusion of a plant 5-methylcytosine DNA glycosylase to a sequence-specific DNA binding domain

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