Tracking-seq reveals the heterogeneity of off-target effects in CRISPR-Cas9-mediated genome editing.
Nat Biotechnol
; 2024 Jul 02.
Article
in En
| MEDLINE
| ID: mdl-38956324
ABSTRACT
The continued development of novel genome editors calls for a universal method to analyze their off-target effects. Here we describe a versatile method, called Tracking-seq, for in situ identification of off-target effects that is broadly applicable to common genome-editing tools, including Cas9, base editors and prime editors. Through tracking replication protein A (RPA)-bound single-stranded DNA followed by strand-specific library construction, Tracking-seq requires a low cell input and is suitable for in vitro, ex vivo and in vivo genome editing, providing a sensitive and practical genome-wide approach for off-target detection in various scenarios. We show, using the same guide RNA, that Tracking-seq detects heterogeneity in off-target effects between different editor modalities and between different cell types, underscoring the necessity of direct measurement in the original system.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Language:
En
Journal:
Nat Biotechnol
/
Nat. biotechnol
/
Nature biotechnology
Year:
2024
Document type:
Article