Small CRISPR RNAs Guide Antiviral Defense in Prokaryotes

By Stan J. J. Brouns, M. Jore, Magnus Lundgren, E. Westra, R. Slijkhuis, A. Snijders, M. Dickman, Kira S. Makarova, Eugene V. Koonin, J. van der Oost. Science, 2008

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Prokaryotes acquire virus resistance by integrating short fragments of viral nucleic acid into clusters of regularly interspaced short palindromic repeats (CRISPRs). Here we show how virus-derived sequences contained in CRISPRs are used by CRISPR-associated (Cas) proteins from the host to mediate an antiviral response that counteracts infection. After transcription of the CRISPR, a complex of Cas proteins termed Cascade cleaves a CRISPR RNA precursor in each repeat and retains the cleavage products containing the virus-derived sequence. Assisted by the helicase Cas3, these mature CRISPR RNAs then serve as small guide RNAs that enable Cascade to interfere with virus proliferation. Our results demonstrate that the formation of mature guide RNAs by the CRISPR RNA endonuclease subunit of Cascade is a mechanistic requirement for antiviral defense.

Subfields: Cardiology and Cardiovascular Medicine, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/1dcfc3f2b8750e6781aa9fe7bec62030476dec85

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In Cardiology and Cardiovascular Medicine

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Citations in the corpus

Regulatory RNAs in Bacteria (2009, background): “Until recently, it was not known whether the crRNAs would target DNA or RNA, but CRISPR spacers generated from both strands of phage genes can effectively confer phage resistance (Barrangou et al., 2007; Brouns et al., 2008).”
Development and Applications of CRISPR-Cas9 for Genome Engineering (2014, neutral): “By studying the type I CRISPR locus of Escherichia coli, van der Oost and colleagues showed that CRISPR arrays are transcribed and converted into small crRNAs containing individual spacers to guide Cas nuclease activity (Brouns et al., 2008).”
Crystal structure of Cpf1 in complex with guide RNA and target DNA (2016, background): “…on the architecture of the interference module (Makarova et al., 2015): class 1 systems use a complex of several Cas proteins, as exemplified by Cascade (Brouns et al., 2008; Redding et al., 2015), and class 2 systems use a single enzyme, such as Cas9 (Jinek et al., 2012; Gasiunas et al., 2012).”
CRISPR-Cas systems in bacteria and archaea: versatile small RNAs for adaptive defense and regulation. (2011, background): “coli are described in later sections (14) (57).”
Pseudomonas aeruginosa Lifestyle: A Paradigm for Adaptation, Survival, and Persistence (2017, background): “…which results in the initiation of the cleavage of the crRNA–foreign DNA hybridization complex, mediating survival of bacteria after viral infections while protecting themselves from lysis (Brouns et al., 2008; Mojica et al., 2009; Deveau et al., 2010; Garneau et al., 2010; Bhaya et al., 2011).”
The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli (2011, background): “First, the unique SapI restriction endonuclease site was removed from pCRISPR3 plasmid by XbaI-SapI cleavage followed by re-ligation with T4 DNA ligase to generate pCRISPR3-ΔSapI plasmid.”
Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence (2011, background): “The cells also contained plasmids expressing cas genes (17) and an F-factor encoding pili that are required for M13 adsorption.”
CRISPR/Cas9 systems have off-target activity with insertions or deletions between target DNA and guide RNA sequences (2014, neutral): “Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins constitute a bacterial defense system that cleaves invading foreign nucleic acids (1–8).”
Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System (2013, background): “…RNAs 220 Cell Reports 4, 220–228, July 11, 2013 ª2013 The Authors (crRNAs) with the target DNA sequence, and this requires a third component, the trans-activating CRISPR RNA (tracrRNA), which recruits the crRNA into the Cas9 complex (Brouns et al., 2008; Gasiunas et al., 2012; Jinek et al., 2012).”
CRISPR/Cas system and its role in phage-bacteria interactions. (2010, background): “The Cas proteins are also necessary for the processing of the long CRISPR transcript (14, 19).”
Proteins and DNA elements essential for the CRISPR adaptation process in Escherichia coli (2012, background): “strain lacking casABCDE genes (3,17), and in K-12”
The Biology of CRISPR-Cas: Backward and Forward. (2018, positive): “Cascade of the type I-E CRISPR-Cas system has a molecular weight of 405 kDa and displays the following composition: (Brouns et al., 2008; Jore et al., 2011).”