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High-Resolution Structure of Cas13b and Biochemical Characterization of RNA Targeting and Cleavage.
Slaymaker, Ian M; Mesa, Pablo; Kellner, Max J; Kannan, Soumya; Brignole, Edward; Koob, Jeremy; Feliciano, Patricia R; Stella, Stefano; Abudayyeh, Omar O; Gootenberg, Jonathan S; Strecker, Jonathan; Montoya, Guillermo; Zhang, Feng.
Affiliation
  • Slaymaker IM; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological
  • Mesa P; Protein Structure & Function Programme, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, Structural Molecular Biology Group, University of Copenhagen, Copenhagen, Denmark.
  • Kellner MJ; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Department for Biochemistry and Cell Biology, University of Vienna, Dr Bohr-Gasse 9, 1030 Vienna, Austria.
  • Kannan S; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological
  • Brignole E; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
  • Koob J; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
  • Feliciano PR; Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
  • Stella S; Protein Structure & Function Programme, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, Structural Molecular Biology Group, University of Copenhagen, Copenhagen, Denmark.
  • Abudayyeh OO; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological
  • Gootenberg JS; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological
  • Strecker J; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological
  • Montoya G; Protein Structure & Function Programme, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, Structural Molecular Biology Group, University of Copenhagen, Copenhagen, Denmark.
  • Zhang F; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Department of Biological
Cell Rep ; 26(13): 3741-3751.e5, 2019 03 26.
Article in En | MEDLINE | ID: mdl-30917325
Type VI CRISPR-Cas systems contain programmable single-effector RNA-guided RNases, including Cas13b, one of the four known family members. Cas13b, which has been used for both RNA editing and nucleic acid detection, is unique among type VI CRISPR effectors in its linear domain architecture and CRISPR RNA (crRNA) structure. Here, we report the crystal structure of Prevotella buccae Cas13b (PbuCas13b) bound to crRNA at 1.65 Å resolution. This structure, combined with biochemical experiments assaying the stability, kinetics, and function of Cas13b, provides a mechanistic model for Cas13b target RNA recognition and identifies features responsible for target and cleavage specificity. Based on these observations, we generated Cas13b variants with altered cleavage preferences, which may expand the utility of nuclease-based RNA detection assays and other applications of Cas13b in mammalian cells.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Bacterial Proteins / RNA / Prevotella / Endonucleases / CRISPR-Cas Systems Language: En Journal: Cell Rep Year: 2019 Type: Article
Fulltext
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Bacterial Proteins / RNA / Prevotella / Endonucleases / CRISPR-Cas Systems Language: En Journal: Cell Rep Year: 2019 Type: Article