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In Vivo Selection of mRNA Repair Ribozymes from Randomized Pools
Author(s) -
Müller Ulrich F,
Olson Karen E.
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.882.3
Subject(s) - ribozyme , rna splicing , ligase ribozyme , messenger rna , intron , in vivo , biology , mammalian cpeb3 ribozyme , rna , computational biology , exon , alternative splicing , gene , microbiology and biotechnology , genetics
Group I introns are catalytic RNAs (ribozymes) that self‐splice from pre‐mRNA. These cis‐splicing ribozymes can be converted for gene therapeutic purposes into trans‐splicing ribozymes, which replace the (mutated) 3′‐terminus of an mRNA by the 3′‐exon of the ribozyme, thereby repairing the mRNA. The major limiting step for this strategy is a low in vivo mRNA repair efficiency. We report here an in vivo selection that improves the in vivo mRNA repair efficiency, from millions of ribozyme variants. The improved portions on the ribozymes are the extended guide sequences (EGSs). The predicted secondary structures of the resulting EGSs form several classes of secondary structure motifs, some of which have been found before. The newly discovered motifs could become important for evading cellular responses in therapeutic applications. The absence of a correlation between in vitro and in vivo activities indicates that in vivo methods are most promising for future, stepwise optimizations of mRNA repair ribozymes. This work was supported by the NIH [T32DK007233 to K.E.O].