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Reduction in Gene Expression Noise by Targeted Decondensation of Gene Loci

ABSTRACT: ATAC-seq was done on various cell lines generated from HeLa cells. The cell lines contained dCas9 constructs conjugated to the histone acetyltransferase p300, as well as a similar system with multiple p300 fragments present, per dCas9 enzyme utilizing SunTag.

ORGANISM(S): Homo sapiens

PROVIDER: GSE157399 | GEO | 2020/09/04

REPOSITORIES: GEO

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Site-specific manipulation of Arabidopsis loci using CRISPR-Cas9 SunTag systems

Project description:Understanding genomic functions requires site-specific manipulation of loci via efficient protein effector targeting systems. However, few approaches for targeted manipulation of the epigenome are available in plants. Here, we adapt the dCas9-SunTag system to engineer targeted gene activation and DNA methylation in Arabidopsis. We demonstrate that a dCas9-SunTag system utilizing the transcriptional activator VP64 drives robust and specific activation of several loci, including protein coding genes and transposable elements, in diverse chromatin contexts. In addition, we present a CRISPR-based methylation targeting system for plants, utilizing a SunTag system with the catalytic domain of the Nicotiana tabacum DRM methyltransferase, which efficiently targets DNA methylation to specific loci, including the FWA promoter, triggering a developmental phenotype, and the SUPERMAN promoter. These SunTag systems represent valuable tools for the site-specific manipulation of plant epigenomes.

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DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A [BS-Seq]

Project description:Here, we demonstrate that dCas9-SunTag-DNMT3A dramatically increased CpG methylation at the HOXA5 locus in human embryonic kidney 293T cells (HEK293T). Furthermore, using a single sgRNA, dCas9-SunTag-DNMT3A was able to methylate a 4.5 kb genomic region and repress HOXA5 gene expression. Reduced representation bisulfite sequencing (RRBS) and RNA-seq showed that dCas9-SunTag-DNMT3A methylated regions of interest with minimal impact on the global DNA methylome and transcriptome.

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Reduction in Gene Expression Noise by Targeted Decondensation of Gene Loci

Project description:Reduction in Gene Expression Noise by Targeted Decondensation of Gene Loci

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