Journal
CELL
Volume 165, Issue 2, Pages 488-496Publisher
CELL PRESS
DOI: 10.1016/j.cell.2016.02.054
Keywords
-
Categories
Funding
- NIH [HG004659, NS075449]
- California Institute of Regenerative Medicine [RB3-05009, RB4-06045]
- National Science Foundation Graduate Research Fellowship [DGE-1144086]
- Powell-Focht graduate fellowship
- CJ Martin fellowship from the National Health and Medical Research Council (Australia)
- Division Of Computer and Network Systems
- Direct For Computer & Info Scie & Enginr [1322093] Funding Source: National Science Foundation
Ask authors/readers for more resources
RNA-programmed genome editing using CRISPR/Cas9 from Streptococcus pyogenes has enabled rapid and accessible alteration of specific genomic loci in many organisms. A flexible means to target RNA would allow alteration and imaging of endogenous RNA transcripts analogous to CRISPR/Cas-based genomic tools, but most RNA targeting methods rely on incorporation of exogenous tags. Here, we demonstrate that nuclease-inactive S. pyogenes CRISPR/Cas9 can bind RNA in a nucleic-acid-programmed manner and allow endogenous RNA tracking in living cells. We show that nuclear-localized RNA-targeting Cas9 (RCas9) is exported to the cytoplasm only in the presence of sgRNAs targeting mRNA and observe accumulation of ACTB, CCNA2, and TFRC mRNAs in RNA granules that correlate with fluorescence in situ hybridization. We also demonstrate time-resolved measurements of ACTB mRNA trafficking to stress granules. Our results establish RCas9 as a means to track RNA in living cells in a programmable manner without genetically encoded tags.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available