Circular sisRNA identification and characterisation

Area of Science:

• Molecular Biology
• Genomics
• Non-coding RNA Research

Background:

• Stable Intronic Sequence RNA (sisRNA) are emerging non-coding RNAs with regulatory functions.
• Identifying sisRNA via standard RNA sequencing is challenging due to their low abundance and masking by abundant RNA species.
• sisRNA play roles in gene expression regulation through feedback loops.

Purpose of the Study:

• To present a practical workflow for enriching and detecting circular sisRNA.
• To enable reliable identification of low-abundance sisRNA using deep sequencing.
• To provide methods for validating sisRNA circularity, stability, and function.

Main Methods:

• Utilizing transcriptionally quiescent systems for enrichment.
• Employing ribosomal RNA (rRNA)-depletion strategies.
• Applying RNase R treatment to isolate stable RNA species.
• Deep sequencing for comprehensive identification.

Main Results:

• A workflow enabling reliable detection of circular sisRNA was established.
• The methods facilitate the identification of previously masked sisRNA reads.
• Experimental validation techniques for circularity and stability were presented.

Conclusions:

• The described workflow significantly improves the detection of circular sisRNA.
• This approach aids in understanding the regulatory roles of sisRNA in gene expression.
• Further functional characterization of sisRNA is facilitated by these methods.