Inducible and reversible RNA N⁶-methyladenosine editing.

Area of Science:

• Molecular Biology
• Epigenetics
• Chemical Biology

Background:

• RNA modifications, such as N6-methyladenosine (m6A), play crucial roles in regulating RNA processes and are implicated in human diseases.
• Current methods for studying RNA modifications lack temporal and locus specificity, hindering the understanding of their dynamic functions.
• The need for precise tools to edit RNA modifications is critical for dissecting their context-dependent roles.

Purpose of the Study:

• To develop a novel platform for chemically inducible and reversible RNA m6A modification editing.
• To enable temporal and transcript/locus-specific control over m6A modification dynamics.
• To provide a versatile tool for investigating the functional roles of RNA epigenetic modifications.

Main Methods:

• Integration of chemically induced proximity (CIP) and CRISPR technologies to create an RNA m6A editing platform.
• Utilizing abscisic acid (ABA) as a chemical inducer for temporal control of m6A editing.
• Development of a light-controlled version by incorporating a photo-caged ABA for enhanced spatiotemporal precision.

Main Results:

• Demonstrated temporal control over RNA m6A editing at specific sites within individual transcripts.
• Achieved reversible m6A modification editing through the addition or removal of the ABA inducer.
• Established a photo-switchable system for light-dependent m6A editing, offering advanced spatiotemporal control.

Conclusions:

• The developed platform provides unprecedented temporal and locus-specific control over RNA m6A modifications.
• This system facilitates the dissection of dynamic and context-dependent functions of RNA epigenetic marks.
• The strategy is potentially applicable to the editing of other RNA modifications beyond m6A.