Single Cell Dna Methylation Sequencing

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.

Single cell dna methylation sequencing.

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. 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. Dna methylation is an epigenetic modification that plays an important role in gene expression regulation development and disease. Benefits of methylation sequencing cytosine methylation can significantly modify temporal and spatial gene expression and chromatin remodeling.

However single cell sequencing has been limited in the ability to characterize multiple layers. 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. 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. 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.

Dna methylation chromatin states and their interrelationships represent critical epigenetic information but these are largely unknown in human early embryos. Single cell genome and transcriptome sequencing have revealed ith in several cancer types 8 12. Here we performed single cell dna methylome sequencing for human preimplantation embryos and found that tens of thousands of genomic loci exhibited de novo dna methylation. This finding indicates that genome wide dna methylation reprogramming during preimplantation development is a dynamic balance between strong global demethylation and drastic focused remethylation.

Whole genome bisulphate methylc seq wgbs is the most comprehensive method for dname sequencing providing single base methylation profiles for more than 90 of the cpgs in the genome. Recent technological innovations have spurred the development of methods that enable us to study the occurrence and biology of this mark at the single cell level. 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.