Engineering circular RNA regulators to specifically promote circular RNA production

Area of Science:

• Molecular Biology
• RNA Biology
• Gene Regulation

Background:

• Circular RNAs (circRNAs) are abundant in the mammalian transcriptome and play critical roles in gene regulation and disease development.
• The mechanisms of circRNA biogenesis, regulation, and manipulation are not fully understood.
• Understanding circRNA function requires effective tools for their study and control.

Purpose of the Study:

• To engineer novel regulators that promote circRNA biogenesis.
• To evaluate the efficacy and specificity of these engineered regulators in vitro and in vivo.
• To establish a new method for investigating circRNA biogenesis and manipulating disease-related circRNAs.

Main Methods:

• Engineering circRNA regulators (ECRRs) by combining RNA binding motifs with dimerization domains.
• Utilizing circRNA mini-gene reporters (circGFP, circScreen) to assess ECRR function.
• Employing molecular biology techniques including RT-PCR, qRT-PCR, northern blot, western blot, and flow cytometry.

Main Results:

• ECRRs successfully promoted circRNA production from exogenous reporters in a time- and dose-dependent manner.
• ECRRs demonstrated specificity in enhancing circRNA biogenesis.
• ECRRs were effective in stimulating the production of endogenous circRNAs (circ10720, circBIRC6).

Conclusions:

• The developed ECRRs offer a versatile platform for promoting circRNA biogenesis.
• This approach enables targeted manipulation of circRNA production in vivo.
• The study provides a novel strategy for exploring circRNA functions and their role in diseases.