Fig. 2: IRES-based mRNA vectors with terminal hairpins demonstrate an equivalent eGFP expression as canonical mRNA vectors. | Gene Therapy

Fig. 2: IRES-based mRNA vectors with terminal hairpins demonstrate an equivalent eGFP expression as canonical mRNA vectors.

From: Terminal hairpins improve protein expression in IRES-initiated mRNA in the absence of a cap and polyadenylated tail

Fig. 2: IRES-based mRNA vectors with terminal hairpins demonstrate an equivalent eGFP expression as canonical mRNA vectors.

RNA vector schematics with corresponding eGFP fluorescent signal in transfected HEK293 cells at different time points after electroporation. A mRNA vector schematics for B, C. Vectors -R- and CRA had an eGFP ORF but no IRES. Vectors haRh and ChaRhA had a short poly(A) sequence that flanked the 5′ terminal hairpin (AAAA), an IRES, and the eGFP ORF. Vectors CRA and ChaRhA had a cap (©, ARCA) and a long poly(A) tail (AAAAAA). B eGFP expression when equal amounts of mRNA were used; and C eGFP expression when equal molar concentrations of mRNA were used. D mRNA vector schematics for E. All vectors were identical except where the 5′ cap (C), or long 3′ poly(A) tail (AAAAAA) are indicated. E eGFP fluorescent signal in transfected HEK293 cells 24 and 48 h after electroporation when equal amounts of mRNA were used. Vectors with no IRES are underlined. [h: hairpin; a: short internal poly(A) sequences (AAAA); A: long poly(A) tail (AAAAAA); C: Anti-reverse Cap Analog (ARCA); R: main internal segment that includes the eGFP ORF in non-IRES vectors or an EMCV IRES and the eGFP ORF] (n = 4, *P < 0.05 shown for 24 h only).

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