Multiplexable, locus-specific targeting of long RNAs with CRISPR-Display.

Nat Methods
Authors
Keywords
Abstract

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.

Year of Publication
2015
Journal
Nat Methods
Volume
12
Issue
7
Pages
664-70
Date Published
2015 Jul
ISSN
1548-7105
URL
DOI
10.1038/nmeth.3433
PubMed ID
26030444
PubMed Central ID
PMC4821475
Links
Grant list
P01 GM099117 / GM / NIGMS NIH HHS / United States
P01GM099117 / GM / NIGMS NIH HHS / United States