HDV-like Rz structure contributes to RNA template 5′ module function. (A) Predicted secondary structures of RNA template 5′ modules Rz-28(28) (left) and SL-28 (right). rRNA sequence (blue) and Rz regions mutated across the 5′ modules in this figure (pink) are indicated. (B, left) Analysis of Rz-28(28) cleavage from a PP7 hp or L8 leader sequence. Reaction products were separated by 12% Urea-PAGE. (Right) Depiction of the compared Rz leader sequences described in the text. (C) Quantification of insertion efficiency of TriCasA Rz, SL-28, and Rz-28(28) 5′ modules as determined by flow cytometry. Different Rz-28(28) leader sequences and mutated variants are indicated. Values indicate mean ± SD, n = 3. (D) PCR amplification of 3′ (top) and 5′ (bottom) rDNA-transgene junctions from gDNA in a representative replicate of the experiment in C.
