Project description:Genomic safe harbors (GSHs) are utilized as an ideal integration site for generating transgenic organisms and cells. Discovery of GSHs is one of the crucial factors for the advancement of basic and applied biology in the species. Such GSHs were discovered in Pv11 (Polypedilum vanderplanki) cell line, which can survive extreme desiccation. To identify the integration sites, high-molecular-weight genomic DNAs were extracted. The DNA libraries were prepared and sequenced with a Nanopore MinION sequencer. In the way to confirm that GSHs loci are localized in open chromatin regions we prepared ATAC-seq libraries, which were sequenced in Illumina HiSeq2500.
Project description:Bulk RNA-sequencing experiments were performed to analyze the transcriptomic effects of such integrations into two newly established genomic safe harbor sites. Jurkat and HEK293T cells were edited to integrate CMV-mRuby expressing cassette into Rogi2 genomic safe harbor site using Cas9 RNP
Project description:The CpG depleted Mycoplasma penetrans harbors a CpG specific C5 methyltransferase. The aim of this experiment was to confirm the specificity of the methyltransferase in vivo and in vitro. Genomic DNA from Mycoplasma penetrans and Escherichia coli genomic DNA that either was or was not methylated in vitro by M.MpeI were subjected to Illumina MiSeq bisulfite sequencing.
Project description:Since targeting of specific pathogenic pathways may be more efficient than current unspecific heart failure treatment, we obtained genomewide expression profiles of a DCM subtype characterized by cardiac inflammation (DCMi) in association with parvovirus B19. This study was entirely based on RNA isolated from endomyocardial biopsies so far only rarely used for genomic expression profiling. Keywords: disease state analysis
Project description:Genomic Locus Proteomics of hTERT and MYC promoters in HEK293T cells. dCAS9-APEX2 mediated biotinylation at predefined genomic loci in living cells.
Project description:Human iPSCs and NSCs were engineered by AAVS1 and/or C13 safe-harbor TALENs which mediated targeted integration of various reporter genes at single or dual safe-harbor loci. Multiple clones of targeted human iPSCs were used to compare with parental untargeted NCRM5 iPSCs. Polyclonal targeted human NSCs were used to compare with their parental untargeted NCRM1NSCs or H9NSCs. Total RNA obtained from targeted human iPSCs or NSCs compared to untargeted control iPSCs or NSCs.
Project description:CRISPR-Cas12a-integrated transgenes in genomic safe-harbors retain high expression in human hematopoietic iPSC-derived lineages and primary cells
