Targeting a pre-mRNA structure with bipartite antisense molecules modulates tau alternative splicing

Area of Science:

• Molecular Biology
• Genetics
• RNA Biology

Background:

• Alternative pre-mRNA splicing dysregulation contributes to approximately 15% of human genetic diseases.
• Current antisense molecules typically target continuous RNA sequences, limiting their application for structured RNA elements.

Purpose of the Study:

• To investigate the efficacy of bipartite antisense molecules in targeting pre-mRNA secondary structures.
• To modulate alternative splicing events regulated by RNA structural features, specifically targeting tau pre-mRNA.

Main Methods:

• Design and application of bipartite antisense molecules targeting a tau pre-mRNA hairpin structure.
• Electrophoretic mobility shift assays (EMSA) and RNase protection assays to confirm molecule binding.
• In vitro splicing assays and cell culture models to evaluate the impact on exon 10 splicing.

Main Results:

• Bipartite antisense molecules successfully bind to the targeted tau pre-mRNA hairpin structure.
• These molecules inhibit exon 10 splicing and reverse the effects of disease-associated mutations.
• Demonstrated efficacy in both in vitro splicing assays and cell culture experiments.

Conclusions:

• A novel bipartite antisense strategy can effectively target and modulate RNA secondary structures to alter splicing.
• This approach offers a potential therapeutic strategy for genetic diseases caused by splicing dysregulation.
• The strategy is applicable to other splicing events influenced by RNA structural elements.