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. Author manuscript; available in PMC: 2022 Aug 10.
Published in final edited form as: Nat Chem. 2022 Jun 20;14(8):905–913. doi: 10.1038/s41557-022-00972-7

Figure 4. Characterisation of cap-caged mRNA.

Figure 4

a) PAGE (Polyacrylamide gel electrophoresis) analysis of mRNA from in vitro transcription (IVT) with commercial caps (0, AG) or FlashCaps (1, 2). Shown is one representative gel from n=3 independent replicates. b) Yield and capping efficiency of indicated mRNAs from IVT (50 µL) in the presence of FlashCaps. c) Stability of differently capped (0, 1, 2) mRNA against Dcp1/2 before/after irradiation. Samples were either incubated with enzyme (Dcp1/2) or without (none). Shown is one representative gel from n=3 independent replicates. d) In vitro translation of FlashCap-FLuc-mRNA before/after irradiation. Data of n=3 independent experiments are shown as mean values +/- SD. Statistical significance was determined by two-tailed students t-test. Significance levels were defined as p<0.05:*, p<0.01:**, p<0.001:***. P value for 2(+) versus 2(-) is 9.08 × 10−4. P value for 1(+) versus 1(-) is 2.3 × 10−5. e) Same as d) but with modified nucleotides (m1Ψ, m5C). Data of n=3 independent experiments are shown as mean values +/- SD. Statistical significance was determined by two-tailed students t-test. Significance levels were defined as p<0.05:*, p<0.01:**, p<0.001:***. P value for 2(+) versus 2(-) is 2.69 × 10−4. P value for 1(+) versus 1(-) is 4.3 × 10−5. Abbreviations: nt: nucleotides; irr: irradiated (365 nm, 30 s), M = Marker, AG: ApppG cap, FLuc: Firefly Luciferase, RLuc: Renilla Luciferase, GLuc: Gaussia Luciferase, eGFP: enhanced green fluorescent protein.