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
TLDR
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.Abstract:
RNA has important and diverse roles in biology, but molecular tools to manipulate and measure it are limited. For example, RNA interference can efficiently knockdown RNAs, but it is prone to off-target effects, and visualizing RNAs typically relies on the introduction of exogenous tags. Here we demonstrate 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. After initial screening of 15 orthologues, we identified Cas13a from Leptotrichia wadei (LwaCas13a) as the most effective in an interference assay in Escherichia coli. LwaCas13a can be heterologously expressed in mammalian and plant cells for targeted knockdown of either reporter or endogenous transcripts with comparable levels of knockdown as RNA interference and improved specificity. Catalytically inactive LwaCas13a maintains targeted RNA binding activity, which we leveraged for programmable tracking of transcripts in live cells. Our results establish CRISPR-Cas13a as a flexible platform for studying RNA in mammalian cells and therapeutic development.read more
Citations
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The biogenesis, biology and characterization of circular RNAs.
Lasse Sommer Kristensen,Maria S. Andersen,Lotte V. W. Stagsted,Karoline K. Ebbesen,Thomas B. Hansen,Jørgen Kjems +5 more
TL;DR: Advances in high-throughput RNA sequencing and circRNA-specific computational tools have driven the development of state-of-the-art approaches for their identification, and novel approaches to functional characterization are emerging.
Journal ArticleDOI
Gene regulation by long non-coding RNAs and its biological functions.
TL;DR: A review of the mechanisms of lncRNA biogenesis, localization and functions in transcriptional, post-transcriptional and other modes of gene regulation, and their potential therapeutic applications is presented in this article.
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
The Biogenesis, Functions, and Challenges of Circular RNAs.
TL;DR: The recent progress on circRNA biogenesis and function is surveyed and technical obstacles in circRNA studies are discussed.
Journal ArticleDOI
RNA editing with CRISPR-Cas13
David Benjamin Turitz Cox,Jonathan S. Gootenberg,Omar O. Abudayyeh,Brian Franklin,Max J. Kellner,Julia Joung,Feng Zhang +6 more
TL;DR: A type VI CRISPR-Cas system containing the programmable single-effector RNA-guided ribonuclease Cas13 is profiled in order to engineer a Cas13 ortholog capable of robust knockdown and REPAIR presents a promising RNA-editing platform with broad applicability for research, therapeutics, and biotechnology.
References
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