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. Author manuscript; available in PMC: 2022 Aug 10.
Published in final edited form as: Nat Biotechnol. 2022 Feb 10;40(6):938–945. doi: 10.1038/s41587-021-01171-4

Figure 3: In vitro activity of cadRNAs.

Figure 3:

(a) Plasmid delivered in situ cadRNA generation: RNA editing efficiencies across various transcripts observed in HEK293FT and K562 cells via plasmid delivered circular.200.100 adRNA, 48 hours post transfections are shown. Values represent mean +/− SEM (n=3). These experiments were carried out using either cadRNA or cadRNA.loops.interspersed from Figure 2b. Associated changes in expression levels of target transcripts as compared to levels seen in untransfected controls is also shown, 48 hours post transfections (p=0.2599, p=0.0135, p=0.1982, p=0.7871, p=0.0144, p=0.2674, p=0.1168, p=0.7852, p=0.5145; unpaired t-test, two-tailed). (b) In vitro transcribed (IVT) circular adRNA generation: Linear forms of twister ribozyme flanked circular adRNAs were transcribed in vitro using a T7 polymerase, purified using LiCl, and transfected into cells, where they circularize in situ by the endogenous RNA ligase RtcB. RNA editing efficiencies across various transcripts observed in HEK293FT and K562 cells via IVT circular adRNA, 24 hours post transfections are shown. Values represent mean +/− SEM (n=3). Associated levels of IVT and plasmid delivered circular.200.100 adRNA targeting RAB7A measured in transfected HEK293FT cells 24 hours post transfections are also shown. Values represent mean +/− SEM (n=3).