Splicing correction assay

This splicing correction assay has now been widely adopted, since it has a low background and provides a positive read-out with a large dynamic response. Neutral steric-block ON, such as peptide nucleic acids (PNA) (5) or phospho-rodiamidate morpholino oligonucleotides (PMO) (6). are particularly suitable for this purpose, since they cannot recruit RNase H, they hybridize with high affinity and selectivity to complementary RNA, and they are metabolically very stable. However, they cannot be transfected with most commercially available delivery vectors. 

Luciferase Assay of Splicing Correction by PNA and PNA-Peptide Conjugates... 

Fig. 4. Assay of splicing correction Luciferase activity (a) and RT-PCR evaluation (b).

R6Pen (15) or (R-Ahx-R)4 (16) PNA and PMO conjugates allow splicing correction in this assay with submicromolar EC50 values. 

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