학술논문

Controlling CRISPR-Cas9 with ligand-activated and ligand-deactivated sgRNAs.
Document Type
article
Source
Nature communications. 10(1)
Subject
DNA
RNA
Guide
Ligands
Aptamers
Nucleotide
CRISPR-Cas Systems
Gene Editing
CRISPR-Associated Protein 9
Biochemistry and Cell Biology
Physical Sciences
Biological Sciences
Biotechnology
Genetics
Generic health relevance
RNA
Guide
Kinetoplastida
Language
Abstract
The CRISPR-Cas9 system provides the ability to edit, repress, activate, or mark any gene (or DNA element) by pairing of a programmable single guide RNA (sgRNA) with a complementary sequence on the DNA target. Here we present a new method for small-molecule control of CRISPR-Cas9 function through insertion of RNA aptamers into the sgRNA. We show that CRISPR-Cas9-based gene repression (CRISPRi) can be either activated or deactivated in a dose-dependent fashion over a >10-fold dynamic range in response to two different small-molecule ligands. Since our system acts directly on each target-specific sgRNA, it enables new applications that require differential and opposing temporal control of multiple genes.