Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1159749 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR30633442.fastq.gz | Fastqsanger.gz | |||
| SRR30633443.fastq.gz | Fastqsanger.gz | |||
| SRR30633444.fastq.gz | Fastqsanger.gz | |||
| SRR30633445.fastq.gz | Fastqsanger.gz | |||
| SRR30633446.fastq.gz | Fastqsanger.gz |
Items per page: 5 1 - 5 of 5 |
Similar Datasets
Project description:CRISPR-Cas is an RNA-based defense system that enables prokaryotes to recognize invading foreign DNA by cognate crRNA guides and destroy it by CRISPR-associated Cas nucleases 1,2 . Elucidation of the interference mechanism of the Streptococcus pyogenes Type II CRISPR- Cas9 system has allowed for the successful repurposing of SpCas9 as a generic genome editing tool, with great promise for human gene therapy 3 . However, especially for therapeutic applications, some caution seems appropriate, because Cas9 systems from some human pathogens may induce a cytotoxic response via an unknown mechanism 4 . Here we show that when released in human cells, Cas9 nucleases from the pathogenic bacteria Campylobacter jejuni and S. pyogenes have the potential to cause severe DNA damage. In the absence of a CRISPR RNA guide, native Cas9 nucleases from both pathogens enter the host nucleus, where their presence leads to promiscuous double stranded DNA breaks (DSBs) and induction of cell death. DSB induction can be reduced to background levels either by saturation of CjCas9 and SpCas9 with crRNA guides or by inactivating their nuclease activity. Our results demonstrate that guide-free Cas9 of bacterial pathogens might play an important role in pathogenicity. Furthermore, we propose that saturating Cas9 with appropriate guide RNAs is crucial for efficient and safe therapeutic applications.
2022-03-02 | PXD006712 | Pride
Project description:Using MethylC-Seq to provide single-base resolution of DNA methylation status in Ws background wild-type (WT), ape1l-1 (Ws background), arp-1 (Col-0 background) and zdp-1 (Col-0 background) mutants MethylC-Seq: 1 WT and 3 mutants examined, Ws background wild-type (WT), ape1l-1 (Ws background), arp-1 (Col-0 background) and zdp-1 (Col-0 background) mutants
2014-11-21 | E-GEOD-52983 | biostudies-arrayexpress
Project description:In bacteria and archaea, CRISPR loci confer adaptive, sequence-based immunity against viruses and plasmids. CRISPR interference is specified by CRISPR RNAs (crRNAs) that are transcribed and processed from CRISPR spacers and repeats. Pre-crRNA processing is essential for CRISPR interference in all systems studied thus far. Here we examine crRNA biogenesis and CRISPR interference in naturally competent Neisseria spp., including the human pathogen N. meningitidis. Our studies reveal a unique crRNA maturation pathway in which crRNA transcription is driven by promoters that are embedded within each repeat, yielding crRNA 5’ ends are not formed by processing. Although crRNA 3’ end formation occurs through RNase III cleavage of a pre-crRNA/tracrRNA duplex, as in other Type II CRISPR systems, this processing event is dispensable for interference. The meningococcal pathway is the most streamlined CRISPR/cas system characterized to date. Endogenous CRISPR spacers frequently target genomic sequences of other Neisseria strains and so limit natural transformation, which is the primary source of genetic variation that contributes to immune evasion, antibiotic resistance, and virulence in N. meningitidis. dRNA-seq approach for RNA samples from cultures of N. lactamica 020-06, harvested at mid-log. Two cDNA libraries from total RNA were prepared to distinguish between transcripts with either primary orprocessed 5’ ends: one library is generated from untreated RNA, whereas the other is treated with terminator exonuclease (TEX),
2013-04-23 | E-GEOD-44582 | biostudies-arrayexpress
Project description:To investigate selectivity of mRNA oxidation, total RNA and oxidized RNA isolated from Neuro 2a cells before and after H2O2 treatment were employed for microarray analysis. It was found that selective oxidation of mRNA already occurs under normal culture conditions but was increased by H2O2, especially in a subset of mRNAs related to certain functions. Moreover, mRNA oxidation level is also related to its abundance or stability (half-life time). This shows for the first time that mRNA oxidation is associated with RNA homeostasis including function, stability and abundance depending on cellular redox status in a genome-wide scale. Neuro 2a cells received hydrogen peroxide treatment or no treatment as a control. Samples were applied for RNA extraction and ARP labeling, which could bind with apurinic/apyrimidinic sites, and then a pull-down process to isolate oxidized RNA. Total RNA and oxidized RNA were used for subsequent transcriptomic profiling. 4 types of samples were analyzed: Basal-total: untreated N2a cells labeled with ARP, but not processed for the pull-down assay. Ox-total: hydrogen peroxide-treated N2a cells labeled with ARP, but not processed for the pull-down assay. Basal-ARP: untreated N2a cells labeled with ARP, and processed for the pull-down assay. ARP-derivatized RNA, which is also oxidized RNA, was concentrated and used for the microarray analysis. Ox-ARP: hydrogen peroxide-treated N2a cells labeled with ARP, and processed for the pull-down assay. ARP-derivatized RNA, which is also oxidized RNA, was concentrated and used for the microarray analysis.
2012-10-01 | E-GEOD-39641 | biostudies-arrayexpress
Project description:To gain a global view on the impact of the collateral activity on protein expression levels, mass spectrometry was used for proteomics analysis of HEK 293T cells when PspCas13b was co-expressed with non-targeted (NT) crRNA or targeting (T) crRNA, which targets the BCR-ABL1 mRNA breakpoint. There was no evidence of off-target protein degradation by T crRNA compared to the non-targeting crRNA. The only significantly repressed proteins were the target BCR-ABL1 p190 (81% reduction) and eGFP (89% reduction) that are encoded on the same mRNA expression construct and encoded mRNA. Catalytic dead Cas13 (dCas13) and crRNA only were used as negative control.
2024-05-20 | PXD051089 | Pride
Project description:To determine what signalling pathways are affected by LILRB1 in MM cells, ARP-1 MM cell lines were transfected with lentivirus to knockdown LILRB1, injected to nsg mice, sorted from the bone marrow of NSG mice and sent for RNA-seq. Total RNAs of 2 x 10^6 CTR-KD ARP-1 cells or LILRB1-KD ARP-1 cells were extracted by RNeasy Mini Kit (Qiagen). 5-10 µg RNA samples were sent to Cancer Genomics Center at The University of Texas (Houston, TX) for RNA-seq followed by data analysis. We use the RNA-seq data to determine differential expression of genes in CTR-KD ARP-1 cells and LILRB1-KD ARP-1 cells.
2024-03-07 | GSE226821 | GEO
Project description:The associated experiments document the production of small RNA (sRNA) during the expression of Cas13 and crRNA, crRNA alone, or controls from agrobacterium spot infiltration in Nicotiana benthamiana. We document the specific production of sRNA corresponding to the guide sequence of the targeted mRNA. In cases where a multi-guide crRNA or a hairpin were expressed, abundent sRNA are produced correspinding to the target mRNA, but outside of the corresponding guide sequence site.
2021-12-07 | GSE171980 | GEO