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Leveraging the power of next generation sequencing ngs both genome wide analysis and targeted approaches can provide researchers with insight into methylation patterns at a single nucleotide level.
Single cell dna methylation sequencing. It can also be used to establish composite methylomes that account for cell to cell heterogeneity in complex tissue samples. The introduction of single cell dna methylation sequencing dna methylation is recognized as a principal contributor to the normal development and regulation of gene expression. Single cell dna methylome sequencing quantifies dna methylation. Dna methylation is an epigenetic modification that plays an important role in gene expression regulation development and disease.
The described method enables single cell analysis of dna methylation in a broad range of biological systems including embryonic development stem cell differentiation and cancer. Single cell genome and transcriptome sequencing have revealed ith in several cancer types 8 12. Here we apply single cell chromatin overall omic scale landscape sequencing sccool seq to generate a genome wide map of dna methylation and chromatin accessibility at single cell resolution during human preimplantation development. Line joxm 1 an induced pluripotent stem cell ipsc line derived from fibroblasts cells from hipsci project was cultured and triggered into differentiation towards endoderm.
Dna methylation chromatin states and their interrelationships represent critical epigenetic information but these are largely unknown in human early embryos. It is essential for maintenance of cellular identity and is associated with a number of key processes including genomic imprinting x chromosome inactivation repression of transposable elements aging and carcinogenesis. However single cell sequencing has been limited in the ability to characterize multiple layers. Benefits of methylation sequencing cytosine methylation can significantly modify temporal and spatial gene expression and chromatin remodeling.
There are several known types of methylation that occur in nature including 5 methylcytosine 5mc 5 hydroymethylcytosine 5hmc 6 methyladenine 6ma and 4mc 4 methylcytosine 4mc. Single cell transcription and methylation data was generated from a single donor from the human induced pluripotent stem cells initiative hipsci 15 16 using the previously described protocol for single cell methylation and transcriptome sequencing in the same cells scm t seq see for details. This finding indicates that genome wide dna methylation reprogramming during preimplantation development is a dynamic balance between strong global demethylation and drastic focused remethylation.
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