Project description:We performed a large-scale genome-wide characterisation of indels generated following editing with CRISPR/Cas9. We used pools of sgRNAs and performed targeted capture and sequencing of the edited regions in HepG2 cells.
Project description:In order to identify the effects of ATP7B H1069Q overexpression on the liver transcriptome, we performed Affymetrix GeneChip hybridization experiments. Transcriptome analysis of the hepatocytes overexpressing ATP7B H1069Q, compared with hepatocytes overexpressing ATP7B WT. For the analysis on the hepatocytes overexpressing ATP7B H1069Q, total RNA was extracted from HepG2 cells; RNA extracted from hepatocytes overexpressing ATP7B WT was used as control.
Project description:To understand whether hepatic cells use autophagy to defend against Cu toxicity in WD, we employed ATP7B knockout (ATP7B-KO) HepG2 cell line, by Quant-seq experiments. Transcriptome analysis of the of ATP7B KO cells and WT cells treated with Copper (Cu) compared with untreated cells.
Project description:To identify target genes of the histone demethylase Phf8 in oligodendroglial cells in a genome-wide and unbiased approach, RNA-sequencing studies were performed on wildtype CG4 cells and CRISPR/Cas9-edited Phf8 knockout cell lines. We compared three wildtype and one CRISPR/Cas9-treated clone without genome editing with four CRISPR/Cas9 Phf8-knockout clones. Among others we detected substantial changes in the expression of genes associated with cell cycle and replication of DNA.
Project description:This dataset contains ChIP-seq data of H3K4me3 and Pol III in single cell-derived control and CRISPR/Cas9 induced tRNA gene deletion clones in human cancer cell lines HAP1 and HepG2. In this study, we looked into functional Cas9-induced on-target genomic alteration in our tRNA gene deletion clones, HAP1 t72 and HepG2 t15.
Project description:throughput profiling of WT1 in M15 mouse mesonephric cells and M15 genome edited for the KTS isoforms. The genome-edited lines were derived from the M15 cells by transfection and puromycin selection of cells with the guide RNA expressing plasmids. Briefly, the guide RNAs were designed against the splice site that is essential for KTS isoform. Out of the designed guide RNAs, some of them were cloned into the pX601 vector with the Sa Cas9 variant, and clone confirmation was done using sequencing. Isoform-specific lines were established by providing oligos for repair which represented the region corresponding to the codons coding for KTS or not.
Project description:The aim of this study was to elucidate the impact of ATP7B deficiency on hepatic gene expression profiles in mice fed a high-fat diet. Systemic ATP7B-knockout (Atp7b-/-) mice on a C57BL/6J genetic background were generated via CRISPR/Cas9 technology and subjected to a high-fat, high-fructose, and high-cholesterol diet (abbreviated as high-fat diet, HFD) for 24 weeks to establish a steatohepatitis model. Hepatic gene-expression profiles were analyzed by RNA sequencing (RNA-seq). ATP7B deficiency exacerbated hepatic inflammation and fibrosis in HFD-fed mice, as revealed by RNA-seq data.
Project description:As part of a study to understand the dynamics of chromatin looping and its regulation by CTCF and cohesin, we have determined the genome-wide binding profiles of CTCF and cohesin (Smc1a subunit) in a genome-edited mouse Embryonic Stem Cell (mESC) line referred to as clone C65. We have also performed 3D genome mapping using Micro-C in another genome-edited mESC line referred to as clone C36.
Project description:We recreated the t(7;12) translocation in K562 cells by CRISPR/Cas9 to understand its effects on haematopoietic cells, which is of relevance to understand how this cytogenetic abnormalities causes and promotes acute leukaemia in infants. Wild-type K562 were edited by electroporation of ribonucleoprotein complexes consisting of Cas9 enzyme and two guide RNAs targeting patient-specific breakpoint loci. K562 electroporated with Cas9 enzyme only were used as control. Edited K562 harbouring the t(7;12) were single-cell cloned to obtain homogeneous populations (hereby referred to as K562-t(7;12)). We performed RNA sequencing analysis of K562-t(7;12) compared to K562 control to uncover transcriptional changes associated with the translocation.