Titlebook: Epigenome Editing; Methods and Protocol Albert Jeltsch,Marianne G. Rots Book 2018 Springer Science+Business Media, LLC, part of Springer Na

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Allele-Specific Epigenome Editingrect epigenetic editors to virtually any genomic site. This advancement in DNA-targeting technology brings allele-specific epigenome editing into reach, a “super-specific” variation of epigenome editing whose goal is an alteration of chromatin marks at only one selected allele of the target genomic

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Key to Delivery: The (Epi-)genome Editing Vector Toolboxpy. For this, a functional copy of the gene in question together with elements controlling its expression is produced as a vector and introduced ex vivo into the patient’s own cells that subsequently are reinfused. Alternatively, vectors are administered directly in vivo. Although this strategy resu

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CRISPR/dCas9 Switch Systems for Temporal Transcriptional Controlase-deactivated Cas9 (dCas9) provide scientists with tools to modulate transcription of genomic loci at will by targeting transcriptional effector domains. To interrogate the temporal order of events during transcriptional regulation, rapidly inducible CRISPR/dCas9 systems provide previously unmet m

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Delivery of Designer Epigenome Modifiers into Primary Human T Cellsies toward further understanding how gene expression is regulated and opportunities to harness these properties for therapeutic purposes. In contrast to gene knockout strategies, targeted epigenome modifications, such as editing of DNA methylation, can mediate gene expression modulation without chan

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Stable Expression of Epigenome Editors via Viral Delivery and Genomic IntegrationA-binding proteins. However, the delivery of plasmid DNA to express these fusion proteins via conventional transient transfection has certain consequences which need to be considered during the experimental design. Transient transfection achieves peak gene expression between 24 and 96 h post-transfe

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Non-viral Methodology for Efficient Co-transfectionand functions of the genomic elements and epigenome modifications in an endogenous context and as new methods for treatment of diseases. Application of such technologies has drawn attention, however, to the prevailing lack of effective delivery methods. Promising viral and non-viral methods both cur

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Chromatin Immunoprecipitation in Human and Yeast Cellszation of transcription factors and posttranslational modification of histones. DNA and proteins are reversibly and covalently crosslinked using formaldehyde. Then the cells are lysed to release the chromatin. The chromatin is fragmented into smaller sizes either by micrococcal nuclease (MNase) or s

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