Project description:Recent advances in glioblastoma (GBM) studies provide a comprehensive catalog of its genetic aberrations and cellular heterogeneity. However, a solid understanding of genotype-based analysis of cancer pathway dependency and actionable target identification is required to transform GBM treatment into a personalized era. Here, we generated a spectrum of mutant iPSCs harboring frequent GBM mutations with CRISPR/Cas9 and profiled the organoids (LEGO: Laboratory Engineered Glioblastoma Organoid) derived from these iPSCs temporally on transcriptome, methylome, metabolome, lipidome, proteome, and phospho-proteome levels. We found that LEGOs form brain tumors in vivo and recapitulate critical features of human GBM. The multi-omics analysis discovered essential milestones driven by genetic heterogeneity during GBM progressions, such as lineage alteration, methylome rewriting, and metabolome/lipidome reprogramming, in concordance with altered pathway activity and drug response. This study provides a tool and research path to realizing genome-based personalized GBM therapy using novel advanced models.
Project description:We develop a single cell methylome analysis technique based on RRBS, which works robustly for mouse embryonic stem cells (mESCs), sperm, metaphase II oocytes, and zygotes. In total, 36 samples were analyzed, including 8 single mouse embryonic stem cells (mESCs), pooled-5, 10, 20 mESCs, bulk mESCs, 7 single sperms, 10 single pronuclei from 5 individual zygotes, 2 metaphase II oocytes, 2 the first polar bodies and 3 negative controls.
Project description:The following data comes from two separate experiments. In the initial experiment, luminal and basal murine mammary epithelial cells were analyzed by reduced representation bisulfite sequencing (RRBS) to generate methylome profiles. In the second experiment, luminal and basal murine mammary epithelial cells were analyzed by ATAC-seq.
Project description:The contained data consist of Illumina HiSeq 2500 reads generated from restriction endonuclease digested genomic DNA of Oryza sativa ssp. indica used as proof of concept for plant-RRBS methylome profiling by bioinformatics analyses. Five biological repeats for an inbred control line and a derived epiline of the 4th generation where analysed using two restriction endonuclease combination (MspI-DpnII or MspI-ApekI).
Project description:We performed scATAC-seq on primary glioblastoma tissue samples taken at time of initial resection to map the global chromatin profiles of glioblastoma cells and associated non-neoplastic cells
Project description:The integration of DNA methylation and transcriptional state within single cells is of broad interest. Several single-cell dual- and multi-omics approaches have been reported recently that will enable further investigation into cellular heterogeneity including the discovery and in depth study of rare cell populations. Such analyses will continue to provide important mechanistic insights into the regulatory consequences of epigenetic modifications. Last year we reported a new method for profiling the DNA methylome and transcriptome from the same single cell. Here we present details of the protocol and provide guidance on its utility. Our Smart-RRBS protocol entails physically separating mRNA and genomic DNA and combines Smart-Seq2 and Reduced Representation Bisulfite Sequencing (RRBS). It generates paired epigenetic promoter and RNA-expression measurements for a substantial fraction (approximately one-fourth) of protein-coding genes in a typical single cell. It also works for micro-dissected tissue samples comprising hundreds of cells. The protocol, excluding sequencing, takes ~3 days to process up to 192 samples manually
Project description:The integration of DNA methylation and transcriptional state within single cells is of broad interest. Several single-cell dual- and multi-omics approaches have been reported recently that will enable further investigation into cellular heterogeneity including the discovery and in depth study of rare cell populations. Such analyses will continue to provide important mechanistic insights into the regulatory consequences of epigenetic modifications. Last year we reported a new method for profiling the DNA methylome and transcriptome from the same single cell. Here we present details of the protocol and provide guidance on its utility. Our Smart-RRBS protocol entails physically separating mRNA and genomic DNA and combines Smart-Seq2 and Reduced Representation Bisulfite Sequencing (RRBS). It generates paired epigenetic promoter and RNA-expression measurements for a substantial fraction (approximately one-fourth) of protein-coding genes in a typical single cell. It also works for micro-dissected tissue samples comprising hundreds of cells. The protocol, excluding sequencing, takes ~3 days to process up to 192 samples manually