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ABSTRACT: S. pyogenes (SF370) crRNA sequencing

PROVIDER: PRJNA1160039 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30642334.fastq.gz	Fastqsanger.gz
	SRR30642335.fastq.gz	Fastqsanger.gz
	SRR30642336.fastq.gz	Fastqsanger.gz
	SRR30642337.fastq.gz	Fastqsanger.gz
	SRR30642338.fastq.gz	Fastqsanger.gz

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Project description:S. pyogenes (SF370) ARP-crRNA sequencing

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Rgg-dependent transcriptional regulation in SF370 Streptococcus pyogenes strain

Project description:Rgg-dependent transcriptional regulation in SF370 Streptococcus pyogenes strain was analyzed during post-exponential phase of growth Keywords: rgg mutant Microarray analysis was performed using RNA samples isolated from both wild-type SF370 and SF370 rgg mutant strains during post-exponential phase of growth

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Rgg-dependent transcriptional regulation in SF370 Streptococcus pyogenes strain

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Project description:S. pyogenes (Bruno) crRNA sequencing

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Cas9 of bacterial pathogens induces DNA damage and apoptosis of human cells

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.

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Streptococcus pyogenes

Project description:Rgg-dependent transcriptional regulation in SF370 Streptococcus pyogenes strain

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Processing-Independent CRISPR RNAs Limit Natural Transformation in Neisseria meningitidis

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

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