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Related Experiment Video

Updated: Jun 23, 2026

Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli

Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli

Published on: November 30, 2018

Purifying circular RNA by ultrafiltration.

Karen Guillen-Cuevas, Xiaoming Lu, Marc R Birtwistle

Biorxiv : the Preprint Server for Biology

|December 16, 2024

Summary

This summary is machine-generated.

Circular RNA (circRNA) offers enhanced stability for therapeutics. This study demonstrates ultrafiltration effectively purifies circRNA, achieving 86% purity, surpassing traditional methods for scalable manufacturing.

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Last Updated: Jun 23, 2026

Large-scale Production of Recombinant RNAs on a Circular Scaffold Using a Viroid-derived System in Escherichia coli

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Published on: November 30, 2018

Fecal micro RNA Isolation

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Published on: October 28, 2020

Quantification of Circular RNAs Using Digital Droplet PCR

Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

Area of Science:

Biotechnology
Molecular Biology
Biochemistry

Background:

Messenger RNA (mRNA) vaccines have spurred interest in mRNA therapeutics.
Linear mRNA's short lifespan due to degradation limits its therapeutic potential.
Circular RNA (circRNA) offers superior stability but requires high purity, which is challenging to achieve.

Purpose of the Study:

To explore ultrafiltration for purifying protein-encoding circRNA from in vitro transcription (IVT) mixtures.
To assess the efficacy of ultrafiltration compared to size-exclusion high-performance liquid chromatography (SE-HPLC).

Main Methods:

Investigated ultrafiltration using polyethersulfone membranes (30–300 kDa MWCO).
Measured sieving coefficients for circRNA, linear precursor RNA, and nicked RNA conformers.
Determined critical fluxes and optimal operating conditions for RNA purification.

Main Results:

Achieved 86% purity and >50% yield using ultrafiltration.
Size-exclusion high-performance liquid chromatography (SE-HPLC) yielded 41% purity and 45% yield.
Ultrafiltration demonstrated superior performance in circRNA purification.

Conclusions:

Ultrafiltration is a highly effective method for purifying circRNA at the research scale.
The scalability of ultrafiltration suggests its potential for large-scale circRNA therapeutic manufacturing.
This purification method addresses a key bottleneck in developing stable circRNA-based therapies.