Synthesis and Engineering of Circular RNAs

Area of Science:

• Molecular Biology
• Biochemistry
• Genetics

Background:

• Circular RNAs (circRNAs) are prevalent across life forms, originating from both introns and exons.
• Eukaryotic circRNAs, particularly those derived from exons of protein-coding and noncoding genes, are of significant research interest.
• The demand for synthesized and engineered circRNAs for structure-function studies is high.

Purpose of the Study:

• To present two distinct protocols for the synthesis and circularization of circular RNAs (circRNAs).
• To provide methods for creating circRNAs suitable for structure and function studies in eukaryotic systems.
• To enable the engineering of circRNAs for advanced biological applications.

Main Methods:

• Described are two alternative protocols for RNA synthesis and circularization.
• Methods include chemical synthesis for small, modified RNA segments and enzymatic synthesis for long RNAs.
• Combined strategies of chemical and enzymatic synthesis are employed to generate long, modified linear RNA strands prior to circularization.

Main Results:

• The study details two viable protocols for producing circular RNAs (circRNAs).
• These methods cater to different downstream applications and template structures.
• The protocols facilitate the synthesis of modified RNA strands for subsequent circularization.

Conclusions:

• The developed protocols offer versatile approaches for circRNA synthesis and engineering.
• These methods are essential for advancing the study of circRNA structure and function.
• The ability to synthesize and modify circRNAs is critical for their application in eukaryotic research.