Transcriptome-wide profiling of A-to-I RNA editing by Slic-seq.

Area of Science:

• Molecular Biology
• Genomics
• Biochemistry

Background:

• Adenosine-to-inosine (A-to-I) RNA editing diversifies the transcriptome and influences biological processes.
• Existing methods for identifying RNA editing sites have limitations.

Purpose of the Study:

• To develop and validate a novel method for accurate identification of A-to-I RNA editing sites.
• To investigate the conservation and characteristics of A-to-I editing in human and mouse brains.
• To explore the role of A-to-I editing in neurological diseases.

Main Methods:

• Utilized Endonuclease V for selective cleavage of inosine and sodium periodate for RNA enrichment.
• Applied the method to human brain samples (Alu and non-Alu elements) and various human cell lines.
• Analyzed editing site conservation between human and mouse brains.
• Investigated A-to-I editing in mouse models of Alzheimer's disease, epilepsy, and aging.

Main Results:

• The new method reliably enriches inosine-containing RNA and identifies editing sites.
• Conserved A-to-I editing sites in human cells are predominantly in 3'UTRs, correlating with RNA and protein binding.
• Exonic A-to-I editing shows higher conservation between human and mouse brains compared to other regions.
• A significant decrease in A-to-I editing sites was observed in neuronal activity genes in mouse models of neurological diseases.

Conclusions:

• The developed method provides a robust approach for A-to-I RNA editing site identification.
• A-to-I editing plays a conserved role in gene regulation, particularly in 3'UTRs and exonic regions.
• Dysregulation of A-to-I editing in neuronal activity genes may contribute to neurological disorders.