Association for Molecular Pathology (AMP) 2022 Annual Meeting & Expo

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Why encapsulated RNA (en-RNA)?

Every nucleic acid detection assay requires a reliable, easily quantified positive control. In the case of ribonucleic acid (RNA) testing, stability in the presence of ribonucleases (RNases) is critical. Failure to provide such a control can lead to false negatives and failed experiments. Currently, positive RNA controls often come from in vitro transcription (IVT), inactivated pathogen, and plasmid DNA. Recently, en-RNA has emerged as a leading alternative. Ongoing efforts seek to improve the ease of creating en-RNA controls.

A new system for production of control RNA encapsulated in phage-like particles

Here, we present a single plasmid, double expression system for encapsulating any control RNA sequence within MS2 phage virus coat protein. This technique will produce highly stable, non-infectious capsules of RNA. These capsules make ideal calibration standards or internal assay controls because they are resistant to RNase digestion. Using common extraction methods, it is possible to extract encapsulated template RNA for use as quantification or size standards. Further, en-RNA can be used as a transient gene expression system.

Because our encapsulation technique accommodates RNA sequences of variable length, it is an ideal way to create controls for a variety of applications.