Browse
Submit Data
Databases
API
Help

Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

IPSC RNA-seq upon CRISPRn or CRISPRi manipulation

ABSTRACT: Assess the on- and off-target effects of dox-inducible CRISPR/Cas9 and CRISPRi constructs in a human iPS cell line. Transcript quantification of 3 cell lines, each plus or minus doxycycline and with or without specific single guide RNAs (sgRNAs), with 2 biological replicates each.

ORGANISM(S): Homo sapiens

SUBMITTER: Max Horlbeck

PROVIDER: E-GEOD-76980 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Similar Datasets

iPSC RNA-seq upon CRISPRn or CRISPRi manipulation

Project description:Assess the on- and off-target effects of dox-inducible CRISPR/Cas9 and CRISPRi constructs in a human iPS cell line.

2016-01-19 | GSE76980 | GEO

RNA-seq of CSMD1 enhancer CRISPRi in LUHMES cells and differentiated neurons

Project description:we performed lentiviral CRISPR interference (CRISPRi) by recruiting dCas9 fused with the KRAB domain to the CSMD1 enhancer (fam3) in the neuronal precursor cell line – Lund human mesencephalic (LUHMES). Given that the expression of CSMD1 was not detectable in LUHMES cells we differentiated these cells into neurons. Differentiated neurons with CRISPRi of CSMD1 enhancer showed significantly higher expression of CSMD1 than control.

2022-06-30 | E-MTAB-8027 | biostudies-arrayexpress

Gene expression profiling (RNA-seq) experiment where CRISPRi guides were designed to target the human rs2650492 variant, the promoter of GAPDH, and a non-coding region near TUFM in HEK293t cells

Project description:CRISPRi guides were designed to target genomic regions of interest in HEK293t cells. Four guides were designed per region (see processed file). These regions were an enhancer located in the 3’UTR of SBK1 which harbors the obesity GWAS variant rs2650492, the promoter of GAPDH as a positive control, and a non-coding region near TUFM as a negative control. We also provide transcriptomes for Cas9 transfected cells only. Genes proximal to rs2650492 were assessed for differential expression. This experiment was performed with technical replicates where each replicate is a unique transfection and library preparation.

2021-07-14 | E-MTAB-10460 | biostudies-arrayexpress

Pervasive off-target activity in CRISPR-interference (CRISPRi) system [ChIP-seq]

Project description:CRISPR/Cas system and its dCas derivatives have been widely and effectively used to alter the intended genomic target. However, the presence of off-targets due to unintended binding of sgRNAs to sequences that closely resemble the target sequence, is still a major challenge. Here, we utilized a genome-wide sgRNA library for the dCas9-KRAB CRISPRi system to investigate the presence of off-target activity and its effects on gene expression. Our study provides strong evidence that CRISPRi off-targets affect the transcriptome of a cell extensively. We also highlight that mis-matches to the target DNA are tolerated while sgRNA binding and the length of the contiguous matches in the PAM-proximal region is one of the most predictive features of binding, along with open chromatin and the DNA-RNA hybrid energy. Additionally, we provide a curated random forest model that can be used to predict the off-target activity in the dCas systems.

2024-10-11 | GSE252978 | GEO

Pervasive off-target activity in CRISPR-interference (CRISPRi) system [RNA-seq]

2024-10-11 | GSE252977 | GEO

CHD7 and Runx1 interaction provides a braking mechanism for hematopoietic differentiation

Project description:We investigated the transcriptional consequences of Chd7 and Runx1 loss in 416B cells, a myeloid progenitor cell line. We engineered the cell line to express Cas9 protein and targeted endogenous Chd7 and Runx1 loci with sgRNAs introduced with lentiviral constructs. We read out the transcriptomes of control and perturbed cells using RNA-Seq following an established SMART-Seq2 protocol (Picelli et al. 2014).

2022-04-26 | GSE130719 | GEO

A CRISPRi system for efficient and rapid gene knockdown in Caulobacter crescentus

Project description:CRISPR interference (CRISPRi) is a powerful new tool used in different organisms that provides a fast, specific, and reliable way to knockdown gene expression. Caulobacter crescentus is a well-studied model bacterium, and although a variety of genetic tools have been developed, it currently takes several weeks to delete or deplete individual genes, which significantly limits genetic studies. Here, we optimized a CRISPRi approach to specifically downregulate the expression of genes in C. crescentus. Although the Streptococcus pyogenes CRISPRi system commonly used in other organisms does not work efficiently in Caulobacter, we demonstrate that a catalytically-dead version of Cas9 (dCas9) derived from the type II CRISPR3 module of Streptococcus thermophilus or from Streptococcus pasteurianus can each be effectively used in Caulobacter. We show that these CRISPRi systems can be used to rapidly and inducibly deplete ctrA or gcrA, two essential well-studied genes in Caulobacter, in either asynchronous or synchronized populations of cells. Additionally, we demonstrate the ability to multiplex CRISPRi-based gene knockdowns, opening new possibilities for systematic genetic interaction studies in Caulobacter.

2020-01-23 | GSE139521 | GEO

RNA-seq of CT26 cells expressing control or Gna13 targeting sgRNAs

Project description:CT26 cells expressing lentiviral Cas9 and sgRNAs targeting either control or Gna13

2021-10-01 | E-MTAB-10727 | biostudies-arrayexpress

Discovery of IL2RA and CD69 enhancers using CRISPRa

Project description:We performed tiling CRISPR activation (CRISPRa) screens in Jurkat T cells to discover enhancers controlling IL2RA and CD69 expression. For each target gene, we constructed a pooled lentiviral library of sgRNAs that targeted sites at all Cas9 PAMs throughout the window starting 100 kb upstream of the transcription start site, extending through the gene body, and 25 kb downstream. A separate library was constructed for each gene, and a separate screen was performed for each gene. For each gene, Jurkat cells stably expressing the dCas9-VP64 transcriptional activator were transduced with the lentiviral sgRNA library. Cells were stained for the protein of interested and sorted by flow cytometry into four bins of expression. Distribution of sgRNAs in each of the sorted populations was compared to the unsorted population. Regions where dCas9-VP64 recruitment was sufficient to drive target gene expression were identified as putative enhancers. The hg19 coordinates of the CD69 library window were chr12:9,880,082-10,013,497. The library contained 2,244 negative control sgRNAs taken from the CRISPRi/a libraries described in Gilbert et al., Cell 150, 647-661 (2014). The hg19 coordinates of the IL2RA library window were chr10:6,027,657- 6,204,333. The library contained 2,244 negative control sgRNAs taken from the CRISPRi/a libraries described in Gilbert et al., Cell 150, 647-661 (2014).

2017-04-26 | GSE98178 | GEO

Regulation of Embryonic and Induced Pluripotency by Aurora Kinase-p53 Signaling

Project description:Illumina Beadchip array analyses of mouse embryonic stem cell gene expression profiles in the presence of or upon knockdown of Aurka. An ES complementation 'rescue' system was employed to measure the effects of knockdown (minus Dox). Aurka was rescued to wildtype levels in the presence of Dox (plus Dox). A control rescue ES cell line, with a Luciferase-targeting shRNA, was also assessed.

2012-05-30 | GSE23541 | GEO

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data