Profiling RNA Editing in Single Cells.

Area of Science:

• Molecular Biology
• Genomics
• Epitranscriptomics

Background:

• RNA editing, specifically adenosine to inosine (A-to-I) conversion by ADAR enzymes, is a crucial post-transcriptional modification in humans.
• While studied in tissues, the biological role and dynamics of A-to-I RNA editing within single cells remain underexplored.
• Advancements in single-cell sequencing allow for detailed epitranscriptomic profiling at cellular resolution.

Purpose of the Study:

• To develop and describe a protocol for detecting and characterizing A-to-I RNA editing events in single human pancreatic cells.
• To enable the in-depth investigation of RNA editing's biological role at the single-cell level.
• To leverage publicly available single-cell RNA sequencing data for epitranscriptomic analysis.

Main Methods:

• Utilizing cell sorting techniques combined with deep sequencing.
• Applying a step-by-step protocol to analyze publicly available human single-cell RNA sequencing (scRNA-seq) datasets.
• Focusing on A-to-I editing events in alpha and beta pancreatic cells.

Main Results:

• A detailed protocol is provided for the detection and characterization of A-to-I RNA editing.
• The methodology allows for the analysis of epitranscriptomic dynamics in human pancreatic cells at single-cell resolution.
• Publicly available scRNA-seq data from human alpha and beta cells can be effectively utilized.

Conclusions:

• The described protocol facilitates the study of A-to-I RNA editing in single human pancreatic cells.
• This approach opens new avenues for understanding the functional significance of RNA editing in cellular heterogeneity.
• The methodology is applicable to existing scRNA-seq datasets, promoting further research in epitranscriptomics.