Related Experiment Video
Updated: Aug 6, 2026
09:51
Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
Published on: May 25, 2018
Structure and evolution-guided design of minimal RNA-guided nucleases
Petr Skopintsev1,2, Isabel Esain-Garcia1,2, Evan C DeTurk1,2
1Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
Science (New York, N.Y.)
|July 16, 2026
Summary
Scientists engineered novel RNA-guided nucleases, called SynTnpBs, using AI and structure-guided design. These synthetic genome editors show high activity across diverse cell types, expanding programmable gene editing possibilities.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Engineering
Background:
- Programmable genome editing relies on RNA-guided nucleases.
- Designing novel nucleases beyond natural evolution is challenging but offers expanded capabilities.
- TnpB is a minimal CRISPR-Cas12-like nuclease with potential for engineering.
Purpose of the Study:
- To develop a strategy for designing active, diverse, non-natural RNA-guided nucleases.
- To generate synthetic TnpB variants (SynTnpBs) with enhanced or novel properties.
- To demonstrate the broad applicability and structural basis of the designed nucleases.
Main Methods:
- Utilized a protein design strategy combining structure-guided inverse folding with evolution-informed residue constraints.
- Employed artificial intelligence for generating variant sequences.
- Performed high-throughput screening for functional editors.
- Conducted cryo-electron microscopy for structural determination.
Main Results:
- Successfully generated active and divergent variants of TnpB, termed SynTnpBs.
- SynTnpB editors demonstrated retained or exceeded wild-type activity in bacterial, plant, and human cells.
- Structural analysis revealed stabilizing contacts in RNA-DNA interfaces, confirming design principles.
Conclusions:
- Established a robust strategy for creating non-natural RNA-guided nucleases.
- Demonstrated the potential of AI-driven protein design for expanding the toolkit of genome editing technologies.
- Highlighted the creation of conformationally active nucleic acid binders, broadening the scope of protein design.
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