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

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

doi:10.1038/nmeth.4327

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

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

^*Corresponding author for this work

Obstetrics and Gynecology

Research output: Contribution to journal › Article › peer-review

256 Scopus citations

Abstract

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 â 1/417,000 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.

Original language	English (US)
Pages (from-to)	710-712
Number of pages	3
Journal	Nature Methods
Volume	14
Issue number	7
DOIs	https://doi.org/10.1038/nmeth.4327
State	Published - Jun 29 2017

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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10.1038/nmeth.4327

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abstract = "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 {\^a} 1/417,000 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.",

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

note = "Funding Information: This research was funded through a pilot project (NIDDK P50 DK096373), V Scholar award to M.A. from V Cancer Research Foundation, and UVA Cancer Center pilot project awards to M.A. We would like to thank S. Shang for his constructive feedback and critical reading of the manuscript. Publisher Copyright: {\textcopyright} 2017 Nature America, Inc., part of Springer Nature. All rights reserved.",

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AU - Kuscu, Cem

AU - Parlak, Mahmut

AU - Tufan, Turan

AU - Yang, Jiekun

AU - Szlachta, Karol

AU - Wei, Xiaolong

AU - Mammadov, Rashad

AU - Adli, Mazhar

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

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