Project description:RNA analysis of CRISPR/Cas9 ZNF519 knockout (KO), ZNF441 knockout (KO), ZNF468 knockout (KO) and wild type hESC-derived cortical organoids and ChIP-seq analysis of CRISPR/Cas9 ZNF519 knockout (KO) and wild type hESC-derived cortical organoids, and HEK293 cells with ZNF519 overexpression (OE).
Project description:We used Crispr/Cas9 technology to establish a homozygous clone of EphA2-SE deletion in tumor cells. Wild-type cells (WT) and homozygous cloned cells (EphA2-SE-/-) were selected for high-throughput data detection.
Project description:The goal of this study is to assess the role of ASH1 like histone lysine methyltransferase (ASH1L) in the biology of anaplastic thyroid cancer. CRISPR-Cas9 was used to create 4 independent cell lines derived from BHT-101 anaplastic thyroid cancer cells with premature stop codons prior to the catalytic domain within both alleles of ASH1L. RNA-seq was performed on these 4 KO cell lines, and compared to 3 biological replicates of wild type BHT-101 cells.
Project description:We generated a SNORD71 KO chondrocyte cell pool using CRISPR/Cas9 gene editing. A CRISPR control cell line was generated and used as a control. Levels of 2’-O-methylation of human rRNAs in SNORD71 KO cell pool and CRISPR control cells were evaluated by RiboMethSeq.
Project description:IDH3a KO in NHAs was demonstrated to exert global DNA methylation changes. These studies were followed by an Illumina EPIC methylation array to evaluate the location of differentially methylated CpGs
Project description:Purpose: We idenficied a circRNA, circBNC2, that was downregulated during tubular epithelial cell (TEC) injury. To study the function of circBNC2, we performed CRISPR-Cas9 to knock out circBNC2 in HK2 (a well recognized TEC) cells. Next-generation Sequencing was used to identified the differentially expressed mRNA between circBNC2-KO HK2 cells and wild-type HK2 cells.
Project description:Purpose: We idenficied a circRNA, circBNC2, that was downregulated during hepatocytes injury. To study the function of circBNC2, we performed CRISPR-Cas9 to knock out circBNC2 in L-02 (a well recognized human hepatocytes) cells. Next-generation Sequencing was used to identified the differentially expressed mRNA between circBNC2-KO L-02 cells and wild-type L-02 cells.
Project description:We identified a new RhoE isoform, RhoEα, and compared the differentially regulated signalings by these two proteins. Both RhoE and RhoEα were first knocked out via CRISPR/Cas9 in HEK 293 cells and HeLa cells (KO cells), followed by RhoE and RhoEα reintroduction (RhoE reintroduction cells and RhoEα reintroduction cells, resepectively). Wild-type cells, KO cells, RhoE reintrocution cells, and RhoEα reintroduction cells were then subjected to RNA-seq analysis.
Project description:Adipose Triglyceride Lipase (ATGL) and Monoglyceride Lipase (MGL) are two enzymes that contribute to intracellular neutral lipolysis by breaking down triglycerides stored within lipid droplets. Recently, lipid droplet accumulation has been described as a novel hallmark of cancer. While lipid metabolism has been investigated in cancer in recent decades, the role of lipid hydrolysis and its enzymes have not been in the focus of cancer research. We and others have found that lipid hydrolysis enzymes might play an important role in the development and progression of lung cancer. To this end, we chose four different non-small cell lung cancer cell lines and employed CRISPR-Cas9 gene editing to knock out either ATGL (ATGL-KO) or MGL (MGL-KO), and a non-targeting control (NTC) was employed to generate a control cell line within each parental cell type. We then performed label free quantitative proteomics to identify differences between the generated cell lines and confirmed ATGL-KO in ATGL-KO cell lines as well as MGL-KO in MGL-KO cell lines. Furthermore, dihydroorotate dehydrogenase (DHODH), an enzyme that is important in some cancer, was upregulated in some, but not all, of the NSCLC cancer cell lines lacking either one of the two lipases.
Project description:ANXA4 WT (control) and ANXA4 KO in A498 kidney cancer cells was generated using CRISPR/Cas9 and two independent gRNAs for control and two independent gRNAs targeting ANXA4. Single cells clones (Ctrl.-1, Crtl.-2, KO-1, KO-2) were selected and validated by western blotting confirming loss of ANXA4 in KO cell lines on protein level. Transcriptome profiling (RNA-Sequencing) and differential gene expression analysis of 2 independent replicates per cell line was performed (resulting in 4 control and 4 ANXA4 KO sequencings). ANXA4 KO cells exhibit transcriptional changes compared to WT cells including upregulation of the ETS transcription factor ELF3 and regulation of Matrisome genes.
