Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification

• Published in: Nature biotechnology Vol. 32; no. 6; pp. 577 - 582
• Main Authors: Guilinger, John P, Thompson, David B, Liu, David R
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.06.2014; Nature Publishing Group
• Subjects: 631/1647/1511; Agriculture; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bioinformatics; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Clustered Regularly Interspaced Short Palindromic Repeats; CRISPR-Associated Protein 9; CRISPR-Cas Systems; Deoxyribonucleases, Type II Site-Specific - chemistry; Deoxyribonucleases, Type II Site-Specific - genetics; Deoxyribonucleic acid; DNA; Endonucleases - chemistry; Endonucleases - genetics; Gene Editing - methods; Genetic engineering; Genome, Human; Genomics; Humans; Life Sciences; Protein Multimerization; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; RNA - genetics

A fusion of the FokI nuclease and a catalytically inactive Cas9 is a highly specific genome editing tool. Genome editing by Cas9, which cleaves double-stranded DNA at a sequence programmed by a short single-guide RNA (sgRNA), can result in off-target DNA modification that may be detrimental in some...