Rationally Designed Anti-CRISPR Nucleic Acid Inhibitors of CRISPR-Cas9
| Autor: | Keith T. Gagnon, Masad J. Damha, Kushal J. Rohilla, Daniel O'Reilly, Christopher L. Barkau |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
anti-CRISPR
0301 basic medicine Streptococcus pyogenes Oligonucleotides Computational biology Biochemistry 03 medical and health sciences 0302 clinical medicine Genome editing CRISPR-Associated Protein 9 Drug Discovery Genetics Humans CRISPR Guide RNA Nucleotide Motifs Locked nucleic acid Molecular Biology Gene Editing Chemistry Oligonucleotide Cas9 Original Articles DNA inhibition nucleic acid 3. Good health DNA-Binding Proteins Protospacer adjacent motif 030104 developmental biology Tandem Repeat Sequences 030220 oncology & carcinogenesis Nucleic acid RNA Molecular Medicine CRISPR-Cas9 CRISPR-Cas Systems RNA Guide Kinetoplastida |
| Zdroj: | Nucleic Acid Therapeutics |
| ISSN: | 2159-3345 2159-3337 |
| DOI: | 10.1089/nat.2018.0758 |
| Popis: | Clustered regularly interspaced short palindromic repeat (CRISPR) RNAs and their associated effector (Cas) enzymes are being developed into promising therapeutics to treat disease. However, CRISPR-Cas enzymes might produce unwanted gene editing or dangerous side effects. Drug-like molecules that can inactivate CRISPR-Cas enzymes could help facilitate safer therapeutic development. Based on the requirement of guide RNA and target DNA interaction by Cas enzymes, we rationally designed small nucleic acid-based inhibitors (SNuBs) of Streptococcus pyogenes (Sp) Cas9. Inhibitors were initially designed as 2′-O-methyl-modified oligonucleotides that bound the CRISPR RNA guide sequence (anti-guide) or repeat sequence (anti-tracr), or DNA oligonucleotides that bound the protospacer adjacent motif (PAM)-interaction domain (anti-PAM) of SpCas9. Coupling anti-PAM and anti-tracr modules together was synergistic and resulted in high binding affinity and efficient inhibition of Cas9 DNA cleavage activity. Incorporating 2′F-RNA and locked nucleic acid nucleotides into the anti-tracr module resulted in greater inhibition as well as dose-dependent suppression of gene editing in human cells. CRISPR SNuBs provide a platform for rational design of CRISPR-Cas enzyme inhibitors that should translate to other CRISPR effector enzymes and enable better control over CRISPR-based applications. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|