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

Choongil Lee; Dong Hyun Jo; Gue-Ho Hwang; Jihyeon Yu; Jin Hyoung Kim; Se-Eun Park; Jin-Soo Kim; Jeong Hun Kim; Sangsu Bae

doi:10.1016/j.ymthe.2019.05.013

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

Mol Ther. 2019 Aug 7;27(8):1364-1371. doi: 10.1016/j.ymthe.2019.05.013. Epub 2019 May 24.

Authors

Choongil Lee¹, Dong Hyun Jo², Gue-Ho Hwang³, Jihyeon Yu⁴, Jin Hyoung Kim², Se-Eun Park³, Jin-Soo Kim¹, Jeong Hun Kim⁵, Sangsu Bae⁶

Affiliations

¹ Department of Chemistry, Seoul National University, Seoul 08826, South Korea; Center for Genome Engineering, Institute for Basic Science, Seoul 08826, South Korea.
² Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul 03080, South Korea.
³ Department of Chemistry, Hanyang University, Seoul 04763, South Korea.
⁴ Department of Chemistry, Hanyang University, Seoul 04763, South Korea; Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul 04763, South Korea.
⁵ Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Clinical Research Institute, Seoul National University Hospital, Seoul 03080, South Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, South Korea; Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, South Korea. Electronic address: [email protected].
⁶ Department of Chemistry, Hanyang University, Seoul 04763, South Korea; Research Institute for Convergence of Basic Sciences, Hanyang University, Seoul 04763, South Korea. Electronic address: [email protected].

Abstract

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.

Keywords: CRISPR-Cas9; base editing; nonsense mutation; premature termination codon; stop codon read through.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenine*
Cell Line
Clustered Regularly Interspaced Short Palindromic Repeats*
Codon, Nonsense*
Databases, Genetic
Fibroblasts
Gene Editing*
Genes, Reporter
Humans
Protein Biosynthesis / genetics
RNA, Messenger / genetics

Substances

Codon, Nonsense
RNA, Messenger
Adenine