Fig. 4. Characterization of cap-caged mRNA.
a, PAGE analysis of mRNA from 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 the indicated mRNAs from IVT (50 µl) in the presence of FlashCaps. c, Stability of differently capped (0, 1, 2) mRNA against Dcp1/2 before and 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 and after irradiation. Data of n = 3 independent experiments are shown as mean values ± s.d. Statistical significance was determined by two-tailed Student's t-test. Significance levels were defined as *P < 0.05, **P < 0.01, ***P < 0.001. The P value for 2 (+) versus 2 (−) is 9.08 × 10−4. The 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 ± s.d. Statistical significance was determined by two-tailed Student's t-test. Significance levels were defined as *P < 0.05, **P < 0.01, ***P < 0.001. The P value for 2 (+) versus 2 (−) is 2.69 × 10−4. The P value for 1 (+) versus 1 (−) is 4.3 × 10−5. nt, nucleotide; 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.