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. 2003 Oct;9(10):1254–1263. doi: 10.1261/rna.5450203

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FIGURE 1. — Ribozymes derivatives and their activity in mammalian cells. (A) Variants of the Rib61-3′γ ribozyme shown bound to sickle β-globin target RNA (dashed line). The S0 ribozyme has a 6-nucleotide internal guide sequence (IGS) that is complementary to the target RNA; the S1 ribozyme has a 6-nucleotide IGS and a 41-nucleotide extended guide sequence (EGS) that is complementary to the target RNA; the S3 ribozyme has a 6-nucleotide IGS and a 41-nucleotide nonspecific spacer sequence at its 5′ end; and the S5 ribozyme has a 9-nucleotide IGS, a 35-nucleotide EGS that is complementary to the target RNA, and a 6-nucleotide P10 that is complementary to the 3′exon sequence. (B) The S5Rib61-3′γ ribozyme bound to the sickle β-globin target RNA. (C) Detection of trans-splicing products by RT–PCR amplification. HEK293 cells were transfected with the β^S-globin plasmid (pNL97) alone (lane 8) or in conjunction with plasmids encoding the S5, S1, S3, or S0 ribozymes or an inactive version of the S5 ribozyme (Dead). Total RNA was isolated and trans-splicing products detected by RT–PCR amplification as discussed in Materials and Methods. Amplification of trans-splicing products yield a 116-bp product. Molecular mass markers (MW) of 123 and 110 bp are shown.

FIGURE 1. — Ribozymes derivatives and their activity in mammalian cells. (A) Variants of the Rib61-3′γ ribozyme shown bound to sickle β-globin target RNA (dashed line). The S0 ribozyme has a 6-nucleotide internal guide sequence (IGS) that is complementary to the target RNA; the S1 ribozyme has a 6-nucleotide IGS and a 41-nucleotide extended guide sequence (EGS) that is complementary to the target RNA; the S3 ribozyme has a 6-nucleotide IGS and a 41-nucleotide nonspecific spacer sequence at its 5′ end; and the S5 ribozyme has a 9-nucleotide IGS, a 35-nucleotide EGS that is complementary to the target RNA, and a 6-nucleotide P10 that is complementary to the 3′exon sequence. (B) The S5Rib61-3′γ ribozyme bound to the sickle β-globin target RNA. (C) Detection of trans-splicing products by RT–PCR amplification. HEK293 cells were transfected with the β^S-globin plasmid (pNL97) alone (lane 8) or in conjunction with plasmids encoding the S5, S1, S3, or S0 ribozymes or an inactive version of the S5 ribozyme (Dead). Total RNA was isolated and trans-splicing products detected by RT–PCR amplification as discussed in Materials and Methods. Amplification of trans-splicing products yield a 116-bp product. Molecular mass markers (MW) of 123 and 110 bp are shown.