CRISPR RNA-Dependent Binding and Cleavage of Endogenous RNAs by the Campylobacter jejuni Cas9.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Leenay, Ryan T
Eisenbart, Sara K
Aul, Belinda U
Beisel, Chase L
Sharma, Cynthia M
MetadataShow full item record
AbstractCas9 nucleases naturally utilize CRISPR RNAs (crRNAs) to silence foreign double-stranded DNA. While recent work has shown that some Cas9 nucleases can also target RNA, RNA recognition has required nuclease modifications or accessory factors. Here, we show that the Campylobacter jejuni Cas9 (CjCas9) can bind and cleave complementary endogenous mRNAs in a crRNA-dependent manner. Approximately 100 transcripts co-immunoprecipitated with CjCas9 and generally can be subdivided through their base-pairing potential to the four crRNAs. A subset of these RNAs was cleaved around or within the predicted binding site. Mutational analyses revealed that RNA binding was crRNA and tracrRNA dependent and that target RNA cleavage required the CjCas9 HNH domain. We further observed that RNA cleavage was PAM independent, improved with greater complementarity between the crRNA and the RNA target, and was programmable in vitro. These findings suggest that C. jejuni Cas9 is a promiscuous nuclease that can coordinately target both DNA and RNA.
CitationMol Cell. 2018 Mar 1;69(5):893-905.e7. doi: 10.1016/j.molcel.2018.01.032.
AffiliationHIRI, Helmholtz-Institut für RNA-basierte Infektionsforschung, Josef-Shneider Strasse 2, 97080 Würzburg, Germany.
PublisherElsevier/ Cel Press
The following license files are associated with this item:
- Creative Commons
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 4.0 International
- Programmable RNA Cleavage and Recognition by a Natural CRISPR-Cas9 System from Neisseria meningitidis.
- Authors: Rousseau BA, Hou Z, Gramelspacher MJ, Zhang Y
- Issue date: 2018 Mar 1
- Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems.
- Authors: Yamada M, Watanabe Y, Gootenberg JS, Hirano H, Ran FA, Nakane T, Ishitani R, Zhang F, Nishimasu H, Nureki O
- Issue date: 2017 Mar 16
- The tracrRNA and Cas9 families of type II CRISPR-Cas immunity systems.
- Authors: Chylinski K, Le Rhun A, Charpentier E
- Issue date: 2013 May
- The CRISPR-associated DNA-cleaving enzyme Cpf1 also processes precursor CRISPR RNA.
- Authors: Fonfara I, Richter H, Bratovič M, Le Rhun A, Charpentier E
- Issue date: 2016 Apr 28
- Cut site selection by the two nuclease domains of the Cas9 RNA-guided endonuclease.
- Authors: Chen H, Choi J, Bailey S
- Issue date: 2014 May 9