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Updated: Jan 10, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
Directed evolution of an m6A eraser for site-selective epitranscriptome editing.
Huiqing Zhou1, Chuan-Hui Wang1
1Department of Chemistry, Merkert Chemistry Center, Boston College, Chestnut Hill, MA 02467, USA.
Researchers developed a new method to engineer better RNA demethylase enzymes. This led to FTO-818, an enzyme with 13x higher efficiency, enabling precise control over N6-methyladenosine (m6A) modifications in cells.
Area of Science:
- Molecular Biology
- Biotechnology
- RNA Epigenetics
Background:
- N6-methyladenosine (m6A) is a crucial RNA modification.
- Existing m6A demethylases (FTO, ALKBH5) have limited catalytic efficiency.
- This limitation hinders epitranscriptomic research and biotechnological applications.
Purpose of the Study:
- To develop a platform for enhancing m6A demethylase activity.
- To generate improved m6A demethylase variants for research and biotechnology.
Main Methods:
- Developed a fluorescence-based directed evolution platform for rapid selection.
- Generated and characterized FTO-818, an evolved FTO variant.
- Constructed a dCas13b-FTO-818 fusion for targeted m6A editing.
Main Results:
- Evolved FTO variant (FTO-818) exhibits 13-fold increased catalytic efficiency against m6A.
- FTO-818 shows enhanced demethylation of purified RNAs and endogenous m6A in human cells.
- Site-specific m6A removal achieved using the dCas13b-FTO-818 editor in mRNA and lncRNA.
Conclusions:
- The directed evolution platform successfully generated a hyperactive m6A demethylase.
- FTO-818 is a potent tool for in vitro and in vivo m6A manipulation.
- The dCas13b-FTO-818 editor enables precise, targeted epitranscriptomic editing.
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