Hui Li

RNA targeting with CRISPR–Cas13
Abudayyeh OO, Gootenberg J, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F.
Nature. 2017 Oct 4. doi: 10.1038/nature24049. [Epub ahead of print]
October 4, 2017
Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA. McGovern Institute for Brain Research at MIT, Cambridge, Massachusetts 02139, USA. Department of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA. Departments of Genetics, Biology, and Chemistry, Stanford University, Stanford, California 94305, USA. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455, USA. Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA. Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
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.
Date: 
November 1, 2017
Where: 
HSW 1057 at noon