Journal Article

CRISPR-STOP: gene silencing through base-editing-induced nonsense mutations

Parlak, Mahmut, Mammadov, Rashad, Kuscu, Cem, Szlachta, Karol, Wei, Xiaolong, Yang, Jiekun, Adli, Mazhar, Tufan, Turan

2017

Overview

Early STOP codons created with CRISPR base editors leads to gene knockout with high efficiency and does not stress cells with double-strand DNA breaks. CRISPR-STOP can target the majority of human genes and is useful for genetic screens. CRISPR–Cas9-induced DNA damage may have deleterious effects at high-copy-number genomic regions. Here, we use CRISPR base editors to knock out genes by changing single nucleotides to create stop codons. We show that the CRISPR-STOP method is an efficient and less deleterious alternative to wild-type Cas9 for gene-knockout studies. Early stop codons can be introduced in ∼17,000 human genes. CRISPR-STOP-mediated targeted screening demonstrates comparable efficiency to WT Cas9, which indicates the suitability of our approach for genome-wide functional screenings.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 13/31 / 38/23 / 38/39 / 45/22 / 45/23 / 631/337/572 / 631/61/212 / Bioinformatics / Biological Microscopy / Biological Techniques / Biomedical and Life Sciences / Biomedical Engineering/Biotechnology / brief-communication / Clustered Regularly Interspaced Short Palindromic Repeats / Codon, Nonsense / Codon, Terminator - genetics / Codons / CRISPR / Damage / Deoxyribonucleic acid / DNA / DNA damage / Editing / Gene Expression Regulation / Gene Silencing / Gene targeting / Gene Targeting - methods / Genes / Genetic Vectors / Genomes / Genomics / HEK293 Cells / Humans / Life Sciences / Methods / Mutation / Nucleotides / Physiological aspects / Plasmids / Properties / Proteomics / Transfection