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Related Experiment Video

Updated: Jan 10, 2026

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

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.

Research Square

|November 24, 2025

Summary

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.

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Last Updated: Jan 10, 2026

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In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

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.