Fig. 6: Properties of a selected small replicating RNA species.

a Minimum Free Energy structure of MSRP-22 predicted by RNAfold (default parameters)⁶⁰. Colours indicate sequences aligning to the nucleotides 1–118 within the 5′ UTR (green) and nucleotides 3464–3568 within the 3' UTR (yellow) of MS2wt, respectively. Nucleotides annotated in pink indicate mismatches and grey nucleotides could not be aligned to MS2wt^60,61. Annotation as sense or antisense strand follows the orientation of the matching MS2 genomic or antigenomic strand, respectively. b Sequence alignments of the isolated MS2 RNA parasite MSRP-22 with the 5′ UTR of MS2(+) (top, green) and 3′ UTR of MS2(+) (bottom, yellow), respectively. Alignments were made with Jalview (Version 2.11.1.3)⁶². c Time-course of replication of MSRP-22, analysed by gel electrophoresis on a 2% TAE agarose gel. Reaction mixtures contained varying concentrations of input MSRP-22 antisense RNA (5, 15 or 50 nM, image cropped) with 500 nM MS2rep core complex (top row). Controls contained either no input RNA, no MS2rep or MS2rep with RQ135 RNA as non-specific RNA input (bottom row). The ladder used was RiboRuler Low Range RNA Ladder (Thermo Fisher Scientific).