Project description:MCF-7 is an estrogen receptor-positive breast cancer cell line. This experiment is designed to study (1) the effect of estradiol (E2) exposure and (2) lysine methyltransferase 2B (KMT2B) knockdown in MCF-7 cells. Cells were grown for 72 hours prior to treatment with vehicle or 10 nM E2 for 4 and 24 hours. Additionally, to assess the effect of KMT2B knockdown, MCF-7 cells were transfected with KMT2B targeting siRNA or scrambled control siRNA in the absence or presence of E2. RNA were isolated using Trizol and hybridized to Affymetrix GeneChip Human Genome U133 Plus 2.0 array.
Project description:HBV-KMT2B integrated human induced pluripotent stem cell-derived hepatic progenitor cells (KMT2B-Int HPCs) vs heterozygous mutated KMT2B human induced pluripotent stem cell-derived hepatic progenitor cells (KMT2B-HT HPCs) vs original human induced pluripotent stem cell-derived hepatic progenitor cells (KMT2B-WT HPCs) Transcriptional profiling of all KMT2B-WT, HT, and Int HPCs. KMT2B-HT and Int iPSCs were generated by gene editing using CRISPR/Cas9 and Cre recombinase technologies. iPSCs were differentiated into a hepatic lineage. On Day14 of hepatic differentiation, CD13 high and C133 high hepatic cells were sorted by fluorescence-activated cell sorter (FACS) on feeder mouse embryonic fibroblasts (MEFs). HPCs forms colony on MEFs and were stably cultured with maintaining their hepatic phenotype.
Project description:To investigate the global analysis of H3K4me3 of HBV-KMT2B integrated HPCs, CUT&Tag analysis of HBV-KMT2B integrated HPCs and KMT2B heterozygous knockout HPCs was performed.
Project description:KMT2B,a major H3K4 tri-methyltransferase, contributes to the development of various cancers. However, its role in cervical cancer is unclear. We found that KMT2B is upregulated in cervical cancer tissues, which is significantly associated with poor prognosis.To investigate the underlying mechanism of KMT2B in the progression of cervical cancer, we established stable KMT2B-overexpression HeLa cells.Then we compared the whole genome mRNA expression profile of KMT2B overexpression and control HeLa cells using RNA-seq.
Project description:To investigate the transcriptomic analysis of HBV-KMT2B integrated HCC, RNA-seq analysis of tumor tissue of HBV-KMT2B integrated HCC was performed. To further analyze KMT2B integrations, we identified full-length complementary DNA by long-read RNA-seq (Iso-seq).
Project description:To investigate the transcriptomic analysis of HBV-KMT2B integrated HCC, RNA-seq analysis of tumor tissue of HBV-KMT2B integrated HCC was performed. To further analyze KMT2B integrations, we identified full-length complementary DNA by long-read RNA-seq (Iso-seq).
Project description:HBV-KMT2B integrated human induced pluripotent stem cells (KMT2B-Int iPSCs) vs heterozygous mutated KMT2B iPSCs (KMT2B-HT iPSCs) vs homozygous mutated KMT2B iPSCs (KMT2B-KO iPSCs) vs their original iPSCs (KMT2B-WT iPSCs)
Project description:We identified full-length complementary DNA by long-read RNA-seq (Iso-seq) in HBV-KMT2B integrated HCC sample. To compare the data of HCC sapmle with HBV-KMT2B integrated HPCs, which reproduces the sequence of the HCC sample, we performed long-read RNA-seq in HBV-KMT2B integrated HPCs, as well.
Project description:Transcriptional profiling of all KMT2B-WT, HT, KO, Int iPSCs. KMT2B-HT, KO, and Int iPSCs were generated by gene editing using CRISPR/Cas9 and Cre recombinase technologies.
Project description:The mammalian male germline is sustained by a pool of spermatogonial stem cells (SSCs) that can transmit both genetic and epigenetic information to offspring. However, the mechanisms underlying epigenetic transmission remain unclear. The histone methyltransferase Kmt2b is highly expressed in SSCs and required for the SSC-to-progenitor transition. At the stem cell stage, Kmt2b catalyzes H3K4me3 at bivalent H3K27me3-marked promoters as well as at promoters of a new class of genes lacking H3K27me3, which we call monovalent. Monovalent genes are mainly activated in late spermatogenesis while the bivalent genes are mainly not expressed until embryonic development. These data suggest that SSCs are epigenetically primed by Kmt2b in two distinguishable ways for the upregulation of gene expression during the spermatogenic program and through the male germline into the embryo. Because Kmt2b is also the major H3K4 methyltransferase for bivalent promoters in embryonic stem cells, we also propose that Kmt2b has the capacity to epigenetically prime stem cells.