Development of a ribonuclease containing a G4-specific binding motif for programmable RNA cleavage

• Main Authors: Phan, Anh Tuân, Dang, Dung Thanh
• Other Authors: School of Physical and Mathematical Sciences
• Format: Article
• Language: English
• Published: 2019
• Subjects: DRNTU::Science::Physics; Catalytic RNA; Protein Design
• Online Access: https://hdl.handle.net/10356/106553; http://hdl.handle.net/10220/48945
• Institution: Nanyang Technological University

We developed a ribonuclease for site-specific targeting and cleavage of single-stranded RNA. The engineered RNase protein was constructed by incorporating two independent functional domains, an RNase HI domain that could cleave the RNA strand in a DNA-RNA hybrid, and a domain of the RHAU protein that could selectively recognize a parallel DNA G-quadruplex (G4). The newly designed RNase first recruits a DNA guide oligonucleotide containing both a parallel G4 motif and a template sequence complementary to the target RNA. This RNase:DNA complex targets and efficiently cleaves the single-stranded RNA in a site-specific manner. A major cleavage site occurs at the RNA region that is complementary to the DNA template sequence. The newly designed RNase can serve as a simple tool for RNA manipulation and probing RNA structure.