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Project description:Differential Gene Expression Upon PFN1 Deletion
| PRJNA630391 | ENA
Project description:The histological grade of carcinomas describes the ability of tumor cells to organize differentiated epithelial structures and has prognostic impact. Molecular control of differentiation in normal and cancer cells relies on lineage-determining transcription factors (TFs) that activate the repertoire of cis-regulatory elements controlling cell type-specific transcriptional outputs. TF recruitment to cognate genomic DNA binding sites results in the deposition of histone marks characteristic of enhancers and other cis-regulatory elements. Here we integrated transcriptomics and genome-wide analysis of chromatin marks in human pancreatic ductal adenocarcinoma (PDAC) cells of different grade to identify first, and then experimentally validate the sequence-specific TFs controlling grade-specific gene expression. We identified a core set of TFs with a pervasive binding to the enhancer repertoire characteristic of differentiated PDACs and controlling different modules of the epithelial gene expression program. Defining the regulatory networks that control the maintenance of epithelial differentiation of PDAC cells will help determine the molecular basis of PDAC heterogeneity and progression. Poly(A) fraction of the total RNA from human pancreatic ductal adenocarcinoma cell lines was extracted and subjected to by multiparallel sequencing. Experiments were carried out in unmodified cells in duplicate, genome edited clonal CFPAC1 cells (2 KLF5-deleted CRISPR-Cas9 clones, 3 ELF3-deleted CRISPR-Cas9 clones and 2 wt clones) and CFPAC1 cells ectopically expressing ZEB1 or empty vector control (in duplicate).
2016-01-18 | E-GEOD-64558 | biostudies-arrayexpress
Project description:Purpose: To identify gene expression changes (using RNAsequencing) in MC3T3-E1 cells where Rorβ has been deleted using CRISPR/Cas9 (and a control cell line) Method: MC3T3-Control and MC3T3-Rorβ-CRISPR(KO) cells were osteoblast differentiated for 2 days, and RNA was harvested (n=3 for each cell line) Results: We identified genes and pathways (using IPA) that were significantly altered in the Rorβ-CRISPR(KO) cell line, compared with the Control cell line Conclusions: Many pathways were changed, namely the Wnt pathway which is important to the biology of Rorβ and to osteoblast differentiation in general
2017-12-09 | GSE107849 | GEO
Project description:Human acute myeloid leukemia cell lines OCI-AML2 and OCI-AML3 were used in a CRISPR/Cas9-mediated approach to specifically target DDX3X’s gene sequences encoding the RNA binding domain of the helicase. DDX3X RNA binding domain is bipartite in the two halves of the helicase core. sgRNAs were designed to target both halves of the domain (named RNA binding domain A and B – RBDA and RBDB). We performed RNA-seq to observe the gene expression changes in both OCI-AML2 and OCI-AML3 cell lines following the not-combined CRISPR/Cas9 –mediated targeting of both regions of the DDX3X RNA binding domain. Control CRISPR/Cas9 performed with no sgRNA expressing vector (named “empty vector”) was performed in both cell lines. The latter condition was used as a control for gene expression changes analysis, for each cell line.
2019-11-23 | E-MTAB-6241 | biostudies-arrayexpress
Project description:Genome-wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) associated with increased risk for colorectal cancer (CRC). A molecular understanding of the functional consequences of this genetic variation is complicated because most GWAS SNPs are located in non-coding regions. After identifying H3K27Ac peaks in HCT116 colon cancer cells that harbor SNPs associated with an increased risk for CRC, we used CRISPR/Cas9 nuclease to delete 2 CRC risk-associated H3K27Ac peaks (E7 and E24), a peak that is not associated with CRC risk (18qE) and a region that doesn?t have H3K27Ac peak (18qNE) from HCT116 cells and analyzed effects on the transcriptome and epigenome. We also deleted E7 region from HEK 293 cells and analyzed effects on the transcriptome of HEK 293. We also confirmed the physical interaction between enhancers of our interest and their putative target genes. Analysis of RNA-seq data and ChIP-seq between control clones and enhancer deleted clones in HCT116 cell. For Control, gRNA empty vectorplasmid was transfected with Cas9-GFP. For Deletion, gRNAs that have enhancer target sequences were transfected along with Cas9-GFP. Cells with high GFP expression were identified using fluorescence-activated cell sorting. Sorted cells were plated into individual wells of a 24 well plate and then re-plated as single cells in 10cm dishes and subsequently expanded for further analyses. Samples used in this study were clonal populations. Interaction profiling of enhancers of interest with putative target genes using 4C-seq
2016-01-12 | E-GEOD-72631 | biostudies-arrayexpress
Project description:Three independent parental or NLUCAT1 CRISPR/Cas9-deleted A549 clones were cultured in normoxia (Nx) or exposed to hypoxia (Hx) at 1% O2 for 24 hours.
2019-09-04 | GSE117036 | GEO
Project description:To study the effect of genes that interface inflammatory and metabolic signaling, CRISPR/Cas9 has been used to delete CD5L and RORA in THP-1 human monocytic cell line. Cells transduced with sgRNAs targeting Renilla luciferase gene were used as controls. Deletion and control lines were differentiated with PMA and allowed to rest for 24 hours prior to isolation of total RNA.
2018-12-17 | E-MTAB-7295 | biostudies-arrayexpress
Project description:Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common hereditary cause of kidney failure, resulting from germline mutations in PKD1 and PKD2 in over 90% of cases. Polycystin-1 (PC-1), a cell surface receptor linked to focal adhesion proteins, plays a critical role in regulating cell-matrix and cell-cell interactions. This study explores the hypothesis that the disrupted actin phenotype observed in ADPKD may be influenced by F-actin binding and the translational regulation of actin-associated proteins mediated by a PC1-BICC1 complex. We examined whether actin-associated pathways are deregulated in UCL93 kidney epithelial cells and CRISPR-Cas9 knockout models for PKD1 and BICC1. Total RNA was extracted for gene expression analysis using total RNA-Seq, and the data were utilized to identify deregulated pathways in the knockout models.
2026-07-27 | E-MTAB-14624 | biostudies-arrayexpress
Project description:Cdc42 is a Rho family GTPase known for its central role in cell polarity and cytoskeletal regulation. To understand the role of Cdc42 in polarised secretion from cytotoxic T lymphocytes (CTLs) we used CRISPR/Cas9 gene deletion. While Cdc42-deleted CTLs initially showed reduced secretion for up to two days after CRISPR-mediated deletion, full secretion was rapidly restored and even enhanced while CDC42 protein remained absent. In contrast, chemical inhibition of CDC42 using CASIN consistently decreased secretion in wild-type cells, but had no impact on Cdc42-deleted CTLs, confirming the specificity of this inhibitor. Comparative proteomics and transcriptomics of CTLs after Cdc42-deletion revealed transcriptional changes that could support improved T cell function including compensation via other Rho GTPases. Targeting the promoter region of Cdc42 did not trigger transcriptional adaptation, consistent with a nonsense-mediated decay mechanism of genetic compensation. Our work highlights the importance of taking orthogonal approaches to study protein function, and reveals the remarkable robustness of primary T cells to adapt to loss of an essential gene.
2025-09-08 | PXD063348 | Pride
Project description:STAT3 is determined to play a crucial role in JAK/STAT signaling pathway. The mutations and copy number alteration of STAT3 can cause cancers and other diseases. The 317-567 amino acid domain of STAT3 protein contributes to binding with genomic DNA for gene activation. However, the impact of the mutations within DNA binding domain of STAT3 on the genomic binding pattern is not fully understood. Stat3 was deleted the 400-411 amino acid residue of \\"NNGSLSAEFKHL\\" using CRISPR-CAS9 in human colon cancer cells SW480. The cells were performed STAT3 ChIP-seq to investigate the genome-wide binding pattern of STAT3 between wild type and mutant STAT3 of SW480.
2020-07-06 | E-MTAB-8417 | biostudies-arrayexpress