Effects of CYTOR Knockdown via CRISPRi on K562 cells
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1201158 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:The long noncoding RNA CYTOR has been linked to poor prognosis of cancer, this includes chemo- and radioresistance. Knowing the mechanisms of CYTOR can benefit current cancer therapeuitics. To examine the mechanisms of CYTOR on K562, we performed CRISPRi to knock it down. RNA sequencing were then performed on these knocked down and wild type cells. This is followed by differential gene expression analysis to determine genes and lncRNAs that are differentially regulated when CYTOR is silenced.
2024-12-22 | GSE285084 | GEO
Project description:Stau2 knockdown in human bcCML cells (K562)"
| PRJNA557137 | ENA
Project description:CRISPRi knockdown of essential genes in Pseudomonas aeruginosa
| PRJNA668371 | ENA
Project description:We previous identified enhancers that regulate MYC expression in K562 cells (Fulco et al. Science 2016). Here, we perturbed MYC enhancers with individual CRISPRi gRNAs and performed ChIP-seq to study the effects on chromatin state.
2023-11-14 | GSE225157 | GEO
Project description:We previously characterized zinc finger protein gene HZF1 (ZNF16) and demonstrated its important roles in erythroid and megakaryocytic differentiation of K562 cells by loss-function assay. However its effect in erythroid and megakaryocytic differentiation of hematopoietic stem/progenitor cells (HSPCs) and the mechanisms by which it functions have not been understood. In this study, we detected up-regulation of ZNF16 during erythroid and megakaryocytic differentiation of K562 cells and normal CD34+ HSPCs, and demonstrated that ZNF16 promotes erythroid and megakaryocytic differentiation by gain-of-function and loss-of-function experiments. Gene expression profiling by mRNA array and PCR validation in the K562 transforments with ZNF16 over-expression suggested that cell division cycle-associated 7-like gene (JPO2) and v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog gene (c-KIT) were among the genes regulated possibly by ZNF16. Luciferase reporter assay and Chromatin Immunoprecipitation demonstrated that ZNF16 binds to JPO2 and c-KIT promoters and inhibits their expression in K562 cells. A significant decrease of JPO2 and c-KIT levels was observed during erythroid and megakaryocytic differentiation of K562 and CD34+ cells. The knockdown of either JPO2 or c-KIT partially rescued the differentiation inhibition caused by ZNF16 knockdown. We also found that ZNF16 inhibits c-KIT/c-Raf/MEK/ERK/c-Jun/HEY1 signal pathways, which finally up-regulated expression of GATA1, a central regulator of erthroid and megakaryocyte differentiation. By lentivirus-mediated gene transfer, we demonstrated that enforced expression and knockdown of ZNF16 in HSPCs down-regulated and up-regulated expression of its targets respectively. Our data collectively demonstrate that ZNF16 promotes erythropoiesis and megakaryocytopoiesis via its regulation on JPO2 and c-KIT. 4 samples are analyzed, H4-1 and H4-2 are two stable K562 transductants with ZNF16 over-expression, PC1 and PC2 are stable control K562 transductants. Stable K562 transductants were obtained for RNA extraction and hybridization on an Illumina HumanHT-12 V4.0 expression beadchipe xpression Array platform.
2016-03-29 | E-GEOD-46593 | biostudies-arrayexpress
Project description:ChIP-seq following CRISPRi perturbations to MYC enhancers in K562 cells
| PRJNA934120 | ENA
Project description:CRISPR interference (CRISPRi) genetic screens use programmable repression of gene expression to systematically explore questions in cell biology and genetics. However, wider adoption of CRISPRi screening has been constrained by the large size of single guide RNA (sgRNA) libraries and lack of consensus on the choice of CRISPRi effector proteins. Here, we address these challenges to present next-generation CRISPRi sgRNA libraries and effectors. First, we combine empiric sgRNA selection with a dual sgRNA library design to generate an ultra-compact, highly active CRISPRi sgRNA library. Next, we rigorously compare CRISPRi effectors to show that the recently published Zim3-dCas9 provides an optimal balance between strong on-target knockdown and minimal nonspecific effects on cell growth or the transcriptome. Finally, we engineer a suite of cell lines which stably express Zim3-dCas9 and demonstrate robust on-target knockdown across these cell lines. Our results and publicly available reagents establish best practices for CRISPRi genetic screening.
2022-06-03 | GSE205147 | GEO
Project description:CRISPR-Cas9 knockdown of RBM48 in K562 cells
| PRJNA658070 | ENA
Project description:Knockdown specificity of Mobile-CRISPRi system targeting chromosomal mRFP
| PRJNA556370 | ENA
Project description:The Mobile CRISPRi system with and without mRFP-targeting sgRNA was engineered into Pseudomonas aeruginosa PA14 strain with chromosomally encoded mRFP. RNA was isolated from these strains, and the corresponding cDNA library was synthesized and sequenced in 150 bp paired-end reads. Approximately 1,000,000 reads were collected for each of the two samples, with ~94% alignment to PA14 WT by Bowtie254, and transcripts were counted with HTSeq55. Only genes with a non-normalized read count greater than 1 in both samples were included in analysis, with a coverage of 1286 genes (~20% genome). This data shows that the Mobile CRISPRi system is selective for sgRNA-guided knockdown of mRFP.
2019-07-25 | GSE134771 | GEO