Research theses
This project focuses on the development and optimization of bridge RNA–guided recombinases as next-generation programmable tools for precise DNA manipulation. Derived from IS110 insertion sequence elements, these enzymes utilize a bridge RNA (bRNA) to recognize both target and donor DNA, conferring programmable specificity and enabling a unified mechanism for DNA insertion, inversion, and excision.
This innovative bRNA-mediated recombination system expands the landscape of nucleic acid–guided genome editing technologies beyond CRISPR, opening new directions in synthetic biology and genome engineering.
Objectives
1. Engineer modular bRNA–recombinase constructs to systematically analyze how bRNA architecture influences DNA recognition, binding dynamics, and catalytic efficiency.
2. Apply protein engineering techniques, such as directed evolution, rational mutagenesis, and structure-based computational design, to enhance enzyme activity, thermostability, and target fidelity.
3. Perform functional characterization in both Escherichia coli and mammalian cell models to quantify recombination efficiency, evaluate off-target effects, and establish design parameters for optimal performance.
Impact:
The successful completion of this project will establish a novel and versatile RNA-programmable recombination platform, significantly expanding the molecular toolbox available for reprogramming of cell circuits and therapeutic genome engineering.