KES8-0055
Supramolecularly cooperative dual-targeting strategy to improve specificity of CRISPR-based tools
Topic
S8. Frontiers of Functional Polymers in Biology and Medicine
When and Where
Sep 30, 2026
15:00 - 15:25
Room 108
Session Chairs
Kisuk YANG
Minseok KWAK
Presenter(s)
Dongsheng Liu (The Hong Kong Polytechnic University)
Co-Author(s)
Abstract
CRISPR-based tools, particularly CRISPR/Cas9, have become the leading technology in genetic editing due to their remarkable efficiency and simplicity. However, off-target effects remain a significant challenge that hinders its clinical applications. In this study, we introduce a Dual-Target Editing (DTE) strategy to eliminate off-target effects by expanding the recognition region beyond only guide RNA (gRNA) through supramolecularly cooperative effects and enhanced the specificity. Simply replacing sgRNA by facilely synthesized RNA-DNA chimeras (RDC), the DTE strategy inherited simplicity of CRISPR-based tools in operation, and achieve comparable efficiency in cas9 endonuclease. Validated by high-sensitivity CIRCLE-seq technique, the DTE system can completely eliminate (100%) off-target sites after optimization. Following the same principle, we believe its broad potential across other CRISPR-based tools, such as base editor, Cas12, Cas13, CRISPR interference/activation or other RNA-guided technologies, without additional modifications.
