C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector
Omar O. Abudayyeh,Jonathan S. Gootenberg,Silvana Konermann,Silvana Konermann,Silvana Konermann,Julia Joung,Julia Joung,Julia Joung,Ian Slaymaker,Ian Slaymaker,Ian Slaymaker,David Benjamin Turitz Cox,Sergey Shmakov,Sergey Shmakov,Kira S. Makarova,Ekaterina Semenova,Leonid Minakhin,Konstantin Severinov,Konstantin Severinov,Konstantin Severinov,Aviv Regev,Aviv Regev,Eric S. Lander,Eric S. Lander,Eric S. Lander,Eugene V. Koonin,Feng Zhang +26 more
TLDR
LshC2c2 is a RNA-guided RNase which requires the activity of its two HEPN domains, suggesting previously unidentified mechanisms of RNA targeting and degradation by CRISPR systems.Abstract:
The clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated genes (Cas) adaptive immune system defends microbes against foreign genetic elements via DNA or RNA-DNA interference. We characterize the class 2 type VI CRISPR-Cas effector C2c2 and demonstrate its RNA-guided ribonuclease function. C2c2 from the bacterium Leptotrichia shahii provides interference against RNA phage. In vitro biochemical analysis shows that C2c2 is guided by a single CRISPR RNA and can be programmed to cleave single-stranded RNA targets carrying complementary protospacers. In bacteria, C2c2 can be programmed to knock down specific mRNAs. Cleavage is mediated by catalytic residues in the two conserved Higher Eukaryotes and Prokaryotes Nucleotide-binding (HEPN) domains, mutations of which generate catalytically inactive RNA-binding proteins. These results broaden our understanding of CRISPR-Cas systems and suggest that C2c2 can be used to develop new RNA-targeting tools.read more
Citations
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CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity
Janice S. Chen,Enbo Ma,Lucas B. Harrington,Maria Da Costa,Xinran Tian,Joel M. Palefsky,Jennifer A. Doudna +6 more
TL;DR: It is shown that RNA-guided DNA binding unleashes indiscriminate single-stranded DNA cleavage activity by Cas12a that completely degrades ssDNA molecules, which is also a property of other type V CRISPR-Cas12 enzymes.
Journal ArticleDOI
Nucleic acid detection with CRISPR-Cas13a/C2c2
Jonathan S. Gootenberg,Omar O. Abudayyeh,Jeong Wook Lee,Patrick Essletzbichler,Aaron J. Dy,Aaron J. Dy,Julia Joung,Vanessa Verdine,Nina M. Donghia,Nichole M. Daringer,Catherine A. Freije,Catherine A. Freije,Cameron Myhrvold,Cameron Myhrvold,Roby P. Bhattacharyya,Jonathan Livny,Aviv Regev,Aviv Regev,Eugene V. Koonin,Deborah T. Hung,Pardis C. Sabeti,James J. Collins,Feng Zhang +22 more
TL;DR: A Cas13a-based molecular detection platform, termed Specific High-Sensitivity Enzymatic Reporter UnLOCKing (SHERLOCK), is used to detect specific strains of Zika and Dengue virus, distinguish pathogenic bacteria, genotype human DNA, and identify mutations in cell-free tumor DNA.
Journal ArticleDOI
Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6
Jonathan S. Gootenberg,Omar O. Abudayyeh,Max J. Kellner,Julia Joung,James J. Collins,Feng Zhang +5 more
TL;DR: ShERLOCK as discussed by the authors is a platform that combines isothermal preamplification with Cas13 to detect single molecules of RNA or DNA, which can detect Dengue or Zika virus single-stranded RNA and mutations in patient liquid biopsy samples via lateral flow.
Journal ArticleDOI
RNA targeting with CRISPR-Cas13.
Omar O. Abudayyeh,Jonathan S. Gootenberg,Patrick Essletzbichler,Shuo Han,Julia Joung,Joseph J. Belanto,Vanessa Verdine,David Benjamin Turitz Cox,Max J. Kellner,Aviv Regev,Aviv Regev,Eric S. Lander,Eric S. Lander,Eric S. Lander,Daniel F. Voytas,Alice Y. Ting,Feng Zhang +16 more
TL;DR: It is demonstrated that the class 2 type VI RNA-guided RNA-targeting CRISPR–Cas effector Cas13a (previously known as C2c2) can be engineered for mammalian cell RNA knockdown and binding and is established as a flexible platform for studying RNA in mammalian cells and therapeutic development.
Journal ArticleDOI
Evolutionary classification of CRISPR-Cas systems: a burst of class 2 and derived variants.
Kira S. Makarova,Yuri I. Wolf,Jaime Iranzo,Sergey Shmakov,Omer S. Alkhnbashi,Stan J. J. Brouns,Emmanuelle Charpentier,David R. Cheng,Daniel H. Haft,Philippe Horvath,Sylvain Moineau,Francisco J. M. Mojica,David Scott,Shiraz A. Shah,Virginijus Siksnys,Michael P. Terns,Česlovas Venclovas,Malcolm F. White,Alexander F. Yakunin,Alexander F. Yakunin,Winston X. Yan,Feng Zhang,Roger A. Garrett,Rolf Backofen,John van der Oost,Rodolphe Barrangou,Eugene V. Koonin +26 more
TL;DR: An updated evolutionary classification of CRISPR–Cas systems and cas genes is provided, with an emphasis on the major developments that have occurred since the publication of the latest classification, in 2015, which includes 2 classes, 6 types and 33 subtypes.
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