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Updated: Nov 20, 2025

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
Published on: May 15, 2018
Full-Spectrum Targeted Mutagenesis in Plant and Animal Cells
Brian Iaffaldano1, Jakob Reiser1
1Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, US Food and Drug Administration (FDA), Silver Spring, MD 20993, USA.
New hypermutation tools enable targeted DNA sequence diversity in plants and animals, advancing protein engineering and functional studies in higher organisms. These advancements promise broader applications in biological research and clinical settings.
Area of Science:
- Molecular Biology
- Biotechnology
- Genetics
Background:
- Directed evolution is crucial for protein engineering but faces limitations in higher organisms.
- Previous in situ directed evolution in plants and animals was hindered by the inability to introduce targeted DNA sequence diversity.
Purpose of the Study:
- To introduce novel hypermutation tools for generating targeted DNA sequence diversity in plant and animal cells.
- To overcome limitations of traditional directed evolution in multicellular organisms.
Main Methods:
- Development of hypermutation tools that recruit mutagenic proteins to specific DNA loci.
- Utilizing CRISPR-derived hypermutators to alter DNA nucleotides within user-defined regions.
- Employing error-prone DNA polymerases and highly active DNA deaminases for mutagenesis.
Main Results:
- Achieved targeted alteration of all DNA nucleotides within user-defined genomic regions.
- Demonstrated the potential for generating all transition and transversion substitutions within specified genomic windows.
- Enabled in situ directed evolution in relevant cellular contexts for plants and animals.
Conclusions:
- New hypermutation tools significantly advance in situ directed evolution in higher organisms.
- These tools offer a powerful platform for evolving proteins and RNAs with desired properties.
- Expected to benefit biological research and accelerate clinical applications.
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