Peptide nucleic acid-assisted generation of targeted double-stranded DNA breaks with T7 endonuclease I.
Nucleic Acids Res
; 52(6): 3469-3482, 2024 Apr 12.
Article
in En
| MEDLINE
| ID: mdl-38421613
ABSTRACT
Gene-editing technologies have revolutionized biotechnology, but current gene editors suffer from several limitations. Here, we harnessed the power of gamma-modified peptide nucleic acids (γPNAs) to facilitate targeted, specific DNA invasion and used T7 endonuclease I (T7EI) to recognize and cleave the γPNA-invaded DNA. Our data show that T7EI can specifically target PNA-invaded linear and circular DNA to introduce double-strand breaks (DSBs). Our PNA-Guided T7EI (PG-T7EI) technology demonstrates that T7EI can be used as a programmable nuclease capable of generating single or multiple specific DSBs in vitro under a broad range of conditions and could be potentially applied for large-scale genomic manipulation. With no protospacer adjacent motif (PAM) constraints and featuring a compact protein size, our PG-T7EI system will facilitate and expand DNA manipulations both in vitro and in vivo, including cloning, large-fragment DNA assembly, and gene editing, with exciting applications in biotechnology, medicine, agriculture, and synthetic biology.
Full text:
1
Database:
MEDLINE
Main subject:
Peptide Nucleic Acids
/
Deoxyribonuclease I
/
DNA Breaks, Double-Stranded
Language:
En
Year:
2024
Type:
Article