Sequence- and structure-specific RNA processing by a CRISPR endonuclease

By Rachel E. Haurwitz, Martin Jinek, B. Wiedenheft, Kaihong Zhou, Jennifer A. Doudna. Science, 2010

701 citations recorded by the source corpus.

CRISPR Processing Many bacteria and archaea recognize invading viruses and plasmids. Foreign DNA is integrated into so-called clustered regularly interspaced short palindromic repeat (CRISPR) loci, and transcripts from these loci are processed into RNAs that can target the invading DNA or RNA for destruction. To investigate the molecular basis for this processing, Haurwitz et al. (p. 1355) screened CRISPR-associated (Cas) proteins in the opportunistic pathogen Pseudomonas aeruginosa and found they were capable of cleaving the CRISPR transcripts. The crystal structure of Cas4 with the CRISPR RNA transcript revealed how the protein specifically recognized RNA repeats, as well as the mechanism of endonucleolytic cleavage. How a prokaryotic immune system makes small RNAs that target invading nucleic acids. Many bacteria and archaea contain clustered regularly interspaced short palindromic repeats (CRISPRs) that confer resistance to invasive genetic elements. Central to this immune system is the production of CRISPR-derived RNAs (crRNAs) after transcription of the CRISPR locus. Here, we identify the endoribonuclease (Csy4) responsible for CRISPR transcript (pre-crRNA) processing in Pseudomonas aeruginosa. A 1.8 angstrom crystal structure of Csy4 bound to its cognate RNA reveals that Csy4 makes sequence-specific interactions in the major groove of the crRNA repeat stem-loop. Together with electrostatic contacts to the phosphate backbone, these enable Csy4 to bind selectively and cleave pre-crRNAs using phylogenetically conserved serine and histidine residues in the active site. The RNA recognition mechanism identified here explains sequence- and structure-specific processing by a large family of CRISPR-specific endoribonucleases.

Subfields: Genetics, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/20c961a4476ade9cc68209e0e0906bfa8826d3cf

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Genetics

In Molecular Biology

Citations in the corpus

A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation1[OPEN] (2015, methodology): “Different strategies have been explored, such as a self-processing ribozyme system (Gao and Zhao, 2014), a transfer RNA-processing system (Xie et al., 2015), or the Yersinia pestis CRISPR/Cas Subtype protein4 (Csy4) ribonuclease system (Haurwitz et al., 2010; Tsai et al., 2014).”
The Biology of CRISPR-Cas: Backward and Forward. (2018, positive): “The nucleases cleave the RNA directly downstream of the hairpin, yielding mature crRNAs that are composed of a full spacer ﬂanked by a short repeat-derived 5 0 handle and the 3 0 stem loop (Gesner et al., 2011; Haurwitz et al., 2010; Nam et al., 2012; Sashital et al., 2011).”
Biology and Applications of CRISPR Systems: Harnessing Nature's Toolbox for Genome Engineering. (2016, neutral): “thermophilus (Cas6e), and Pseudomonas aeruginosa (Cas6f) remain stably bound to the 30 handle and eventually become part of the Cascade complex (Brendel et al., 2014; Brouns et al., 2008; Gesner et al., 2011; Haurwitz et al., 2010; Sashital et al., 2011).”
Salt resistant crop plants. (2014, methodology): “Another emerging genome engineering tool is the CRISPR/Cas system (short for Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR)/CRISPR-associated (Cas)) [87,88].”