CRISPR-Pass: Gene Rescue of Nonsense Mutations Using Adenine Base Editors

• Published in: Molecular therapy Vol. 27; no. 8; pp. 1364 - 1371
• Main Authors: Lee, Choongil, Hyun Jo, Dong, Hwang, Gue-Ho, Yu, Jihyeon, Kim, Jin Hyoung, Park, Se-eun, Kim, Jin-Soo, Kim, Jeong Hun, Bae, Sangsu
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 07-08-2019; Elsevier Limited; American Society of Gene & Cell Therapy
• Subjects: Adenine; base editing; Cell Line; Clustered Regularly Interspaced Short Palindromic Repeats; Codon, Nonsense; Codons; CRISPR; CRISPR-Cas9; Databases, Genetic; Deoxyribonucleic acid; DNA; Editing; Fibroblasts; Gene Editing; Genes; Genes, Reporter; Genetic disorders; Humans; Immunoglobulins; Mutation; Nonsense mutation; Nucleotide sequence; Original; Plasmids; premature termination codon; Protein biosynthesis; Protein Biosynthesis - genetics; Proteins; RNA, Messenger - genetics; stop codon read through; Translation termination; nonsense mutation; stop codon read through; premature termination codon; base editing; CRISPR-Cas9
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1016/j.ymthe.2019.05.013

A nonsense mutation is a substitutive mutation in a DNA sequence that causes a premature termination during translation and produces stalled proteins, resulting in dysfunction of a gene. Although it usually induces severe genetic disorders, there are no definite methods for inducing read through of premature termination codons (PTCs). Here, we present a targeted tool for bypassing PTCs, named CRISPR-pass, that uses CRISPR-mediated adenine base editors. CRISPR-pass, which should be applicable to 95.5% of clinically significant nonsense mutations in the ClinVar database, rescues protein synthesis in patient-derived fibroblasts, suggesting potential clinical utility.
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Lee et al. showed that CRISPR-pass, based on adenine base editors, would be a targeted tool for bypassing premature termination codons. This system could be applicable to ∼95% of clinically significant nonsense mutations, related to genetic diseases, in the ClinVar database.