Project description:To assess the biological significances of p-hTERT in ALT cells, we used the CRISPR-Cas9 system targeting the hTERT gene and performed RNA-seq to analyze the profile of gene expression in hTERT-CRISPR clones (clone#1 and #2). The subsequent gene ontology (GO) analysis showed enrichment of genes involved in cell cycle process and DNA replication within genes down-regulated in hTERT-CRISPR clones.
Project description:CRISPR screen: U2OS or U2OS p53KO cells expressing Cas9 were transduced with a whole-genome library of CRISPR sgRNAs, then treated with either DMSO or etoposide. Differential sgRNA abundances were calculated for each condition and used to determine the effect of each single gene knockout on fitness and the drug-induced death rate. RNA-Seq: U2OS or U2OS p53KO cells were cultured with either DMSO or etoposide for 48 hours, and then U2OS cells were incubated in this conditioned media for 8 hours. RNA was collected and use to observe differential expression changes between conditions.
Project description:To assess the biological significances of p-hTERT in ALT cells, we used the CRISPR-Cas9 system targeting the hTERT gene and performed genome-wide CRISPR screening, which identified genes in the Fanconi anemia/BRCA pathway as synthetic lethal partners of hTERT.
Project description:Bulk RNAseq analysis was done on single cell clones of U2OS cells either expressing exogeonous CIITA or an empty vector control to identify genes regulated by CIITA expression.
Project description:Whole-genome CRISPR screen of etoposide-treated U2OS and U2OS p53KO cells, and RNAseq of U2OS cells incubated in media conditioned by etoposide-treated U2OS or U2OS p53KO cells.
Project description:We sequenced mRNA from 6 human cell lines stably over-expressed specific gene or empty vector, and searched for differently expressed genes after gene over-expression as compared to empty vector. mRNA levels were compared as following pairs: U2OS+hTERT vs U2OS+vector; U2OS+hTERTmut vs U2OS+vector; VA-13+hTERT vs VA-13+vector; VA-13+hTERTmut vs VA-13+vector.
