Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display
Open Access
- 1 June 2015
- journal article
- research article
- Published by Springer Nature in Nature Methods
- Vol. 12 (7) , 664-670
Abstract
CRISPR-Display is a modular and flexible platform that targets natural or synthetic noncoding RNA domains to specific genomic loci for functional studies and synthetic biology. Noncoding RNAs play diverse roles throughout biology and exhibit broad functional capacity. To investigate and harness these capabilities, we developed clustered regularly interspaced short palindromic repeats (CRISPR)-Display (CRISP-Disp), a targeted localization method that uses Cas9 to deploy large RNA cargos to DNA loci. We demonstrate that functional RNA domains up to at least 4.8 kb long can be inserted in CRISPR guide RNA at multiple points, allowing the construction of Cas9 complexes with protein-binding cassettes, artificial aptamers, pools of random sequences and natural long noncoding RNAs. A unique feature of CRISP-Disp is the multiplexing of distinct functions at multiple targets, limited only by the availability of functional RNA motifs. We anticipate the use of CRISP-Disp for ectopically targeting functional RNAs and ribonucleoprotein (RNP) complexes to genomic loci.Keywords
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