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Programmable RNA methylation and demethylation using PUF RNA binding proteins
Kouki Shinoda1, Akiyo Suda1, Kenko Otonari1
1Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan. imiki@scl.kyoto-u.ac.jp.
Chemical Communications (Cambridge, England)
|January 8, 2020
Summary
Researchers developed new tools to control RNA methylation, a key process in cells. These tools, using PUF proteins fused to FTO or METTL14 enzymes, enable precise control over specific RNA methylation sites.
Area of Science:
- Molecular Biology
- Epigenetics
- RNA Biology
Background:
- N6-methyladenosine (m6A) is a crucial RNA modification.
- Understanding m6A's function requires precise control over its localization.
- Current methods lack specificity for targeted RNA methylation control.
Purpose of the Study:
- To develop novel methods for controlling local RNA methylation.
- To investigate the function of individual m6A sites.
- To create sequence-specific RNA methylation and demethylation tools.
Main Methods:
- Designing fusion proteins combining PUF RNA-binding domains with FTO or METTL14 enzymes.
- Utilizing programmable RNA-binding protein PUF for sequence specificity.
- Employing FTO as an m6A demethylase and METTL14 as an m6A methyltransferase.
Main Results:
- Successfully engineered FTO-PUF fusion proteins for sequence-specific RNA demethylation.
- Successfully engineered METTL14-PUF fusion proteins for sequence-specific RNA methylation.
- Demonstrated the ability to precisely control m6A modification at targeted RNA locations.
Conclusions:
- PUF-based fusion proteins offer a powerful platform for site-specific RNA epigenetic editing.
- These tools advance the study of m6A RNA modifications and their functional roles.
- The developed methods open new avenues for investigating RNA regulation.
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