Abstract
BackgroundEpigenetic modifications, including DNA methylation, play an important role in gene silencing and genome stability. Consequently, epigenetic dysregulation can cause several diseases, such as cancer, obesity, diabetes, autism, and imprinting disorders.ResultsWe validate three methods for the generation of epigenome-edited mice using the dCas9-SunTag and single-chain variable fragment-TET1 catalytic domain. We generate model mice for Silver-Russell syndrome (SRS), an imprinting disorder, by target-specific DNA demethylation in the H19 differentially methylated region. Like SRS patients, these mice show H19 upregulation and Igf2 downregulation, leading to severe intrauterine and postnatal growth retardation.ConclusionThis is the first report of an imprinting disease model animal generated by targeted demethylation of specific loci of the epigenome in fertilized eggs. Epigenome-edited animals are also useful for exploring the causative epimutations in epigenetic diseases.
Highlights
Epigenetic modifications, including DNA methylation, play an important role in gene silencing and genome stability
After 2 days, green fluorescent protein (GFP)-expressing cells were sorted by fluorescentactivated cell sorting (FACS) to isolate vector-expressing cells
Another possibility is that intrauterine growth retardation in Silver-Russell syndrome (SRS) patients may be driven by a placental phenotype impacting the fetus; embryonic stem cells (ESCs)-derived mice complemented by tetraploid embryos would not have this contribution to the phenotype
Summary
Epigenetic modifications, including DNA methylation, play an important role in gene silencing and genome stability. Epigenetic dysregulation can cause several diseases, such as cancer, obesity, diabetes, autism, and imprinting disorders. DNA methylation is a key epigenetic modification that plays an important role in gene silencing and genome stability [1,2,3]. Compared with other epigenetic modifications, DNA methylation is thought to be relatively stable; it is sometimes affected by environmental change and aging, leading to epigenetic diseases such as cancer, obesity, diabetes, autism, and imprinting disorders [4,5,6,7,8,9]. Before the development of epigenome editing, tools for directly demonstrating which epigenetic changes cause disease were not available. Fusion proteins consisting of eukaryotic DNA methyltransferases or hydroxymethylation enzymes and DNAbinding proteins, such as zinc finger proteins [13], transcription activator-like effectors [14, 15], and catalytically inactive Cas (dCas9), based on the clustered regularly interspaced short palindromic repeat (CRISPR)/ CRISPR-associated protein 9 (Cas9) system [16,17,18], have been used to produce targeted DNA modifications
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