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Transcriptome network analysis of Smug1 knock-out cells by using CRISPR/Cas9 system

ABSTRACT: Transcriptome network analysis of Smug1 knock-out cells by using CRISPR/Cas9 system

PROVIDER: PRJNA606327 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11075220_1.fastq.gz	Fastqsanger.gz
	SRR11075220_2.fastq.gz	Fastqsanger.gz
	SRR11075221_1.fastq.gz	Fastqsanger.gz
	SRR11075221_2.fastq.gz	Fastqsanger.gz
	SRR11075222_1.fastq.gz	Fastqsanger.gz

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CRISPR/Cas9 knock down line

Project description:CRISPR/Cas9 knock down line

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Transcriptome network analysis of Smug1 knock-out cells by using CRISPR/Cas9 system

Project description:Purpose: To build a differential transcriptome network in Smug1 knock-out HepG2 hepatocarcinoma cells. Methods: Transcriptome analysis by the RNA-seq via mRNA pull-down. Results: We constructed transcriptome from Smug1 knock-out cells by using RNA-seq analysis. Nucleosome and miRNA related genes were highly enriched in Smug1 KO HepG2 cells. Conclusions: Smug1 regulate the gene expression related with nucleosome assembly and histone function.

2020-06-30 | GSE145210 | GEO

METTL21B is a novel human lysine methyltransferase of translation elongation factor 1A: discovery by CRISPR/Cas9 knock out

Project description:Lysine methylation is widespread on human proteins, however the enzymes that catalyse its addition remain largely unknown. This limits our capacity to study the function and regulation of this modification. Here we report that human METTL21B is a protein methyltransferase, which methylates lysine 165 of eukaryotic translation elongation factor 1A (eEF1A). The CRISPR/Cas9 system was used to knock out putative protein methyltransferases METTL21B and METTL23 in K562 cells. The known eEF1A methyltransferase EEF1AKMT1 was also knocked out as a control. Targeted mass spectrometry revealed the loss of lysine 165 methylation upon knock out of METTL21B, and the expected loss of lysine 79 methylation on knock out of EEF1AKMT1. No loss of eEF1A methylation was seen in the METTL23 knock out. Recombinant METTL21B was then shown to catalyse methylation on lysine 165 in eEF1A1 and eEF1A2 in vitro, confirming it as the methyltransferase responsible for this methylation site. Stable isotope labelling by amino acids in cell culture (SILAC) proteomic analysis revealed dysregulation of processes related to eEF1A function in knock outs of METTL21B and EEF1AKMT1, but specific dysregulation of ribosomal proteins in the knock out of METTL21B. This indicates a specific function for lysine 165 methylation of eEF1A in human. METTL21B is specific to vertebrates, with its target lysine showing similar evolutionary conservation. We suggest METTL21B be renamed eEF1A-KMT3. This is the first study to specifically generate CRISPR/Cas9 knock outs of the genes encoding putative protein methyltransferases, for the purpose of substrate discovery and site mapping. Our approach should prove useful for the discovery of further novel methyltransferases, and more generally for the discovery of sites for other protein-modifying enzymes.

2017-07-03 | PXD005497 | Pride

study CRISPR/Cas9 knock down line by RNA-seq

Project description:validate and study CRISPR/Cas9 knock down line by RNA-seq

2020-01-08 | GSE126350 | GEO

Transcriptome changes in response to dDAVP stimulation in CRISPR/Cas9 mediated Camk2d knock-out mpkCCD cells

Project description:The Ca2+/CaM-dependent protein kinase 2-delta (CAMK2D) has been proposed to be involved in vasopressin signaling in the renal collecting duct, which controls water and salt excretion by the kidney. RNA sequancing and quantitative proteomics analyses identified the expression of multiple CAMK2 isoforms, with CAMK2D being the most abundant in collecting duct cells. To investigate the role of CAMK2D in regulating RNA expression in response to vasopressin signaling, the transcriptome of CRISPR/Cas9-mediated Camk2d knock-out mpkCCD cells was profiled using RNA-Seq in the presence of vasopressin analogue dDAVP.

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Effect of CRISPR-Cas9 mediated knock-out of integrin alpha2 on the transcriptome of DU145 prostate cancer cell grown as a spheroid culture

Project description:CRISPR-cas9 technology was used to knock out alpha2 integrin in DU145 cells. To create two cell lines that could be compared to each other in an appropriate manner, these cells were transfected either with alpha2 integrin cDNA or empty vector. The objective of the study was to find potential alpha2 integrin regulated genes when the cells were grown as spheroids.

2018-08-02 | GSE111507 | GEO

Identification of genes involved in class switch recombination by genome-wide CRISPR/Cas9 knock-out screen

Project description:Identification of genes involved in class switch recombination by genome-wide CRISPR/Cas9 knock-out screen

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study CRISPR/Cas9 knock down line by RNA-seq

Project description:study CRISPR/Cas9 knock down line by RNA-seq

| PRJNA521691 | ENA

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Project description:Long non-coding RNAs (lncRNAs) are molecular switches in cellular differentiation, movement and in the reprogramming of cell states by altering gene expression patterns. However, in endothelial cells their role is not well understood. LINC00607 is an endothelial-enriched lncRNA. In order to uncover the functional relevance of LINC00607, an endothelial lentiCRISPR/Cas9-mediated knock-out was created and subjected to RNA- and ATAC-Seq, which revealed that LINC00607 is important for many pathways, among them VEGF- and TGF-beta-signalling.

2023-01-09 | GSE199877 | GEO

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Project description:A key limitation of the commonly-used CRISPR enzyme S. pyogenes Cas9 is the strict requirement of an NGG protospacer-adjacent motif (PAM) at the target site, which reduces the number of accessible genomic loci. This constraint can be limiting for genome editing applications that require precise Cas9 positioning. Recently, two Cas9 variants with a relaxed PAM requirement (NG) have been developed (xCas9 and Cas9-NG) but their activity has been measured at only a small number of endogenous sites. Here we devised a high-throughput Cas9 pooled competition screen to compare the performance of both PAM-flexible Cas9 variants and wild-type Cas9 at thousands of genomic loci and across 3 modalities (gene knock-out, transcriptional activation and suppression). We show that PAM flexibility comes at a substantial cost of decreased DNA targeting and cutting. Of the PAM-flexible variants, we found that Cas9-NG outperforms xCas9 regardless of genome engineering modality or PAM. Finally, we combined xCas9 mutations with those of Cas9-NG, creating a stronger transcriptional modulator than existing PAM-flexible Cas9 variants.

2020-01-26 | GSE143892 | GEO

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