Conditional control of alternative splicing through light-triggered splice-switching oligonucleotides

Area of Science:

• Molecular Biology
• RNA Biology
• Gene Regulation

Background:

• The spliceosome machinery regulates gene expression by removing introns from pre-mRNAs.
• Splice-switching oligonucleotides (SSOs) offer targeted control over gene expression by correcting aberrant splicing.
• Current SSO methods lack conditional control and precise spatial-temporal resolution.

Purpose of the Study:

• To develop a method for conditional control of SSO activity with high spatial and temporal resolution.
• To enable optochemical switching of SSO function for precise regulation of alternative splicing.

Main Methods:

• Synthetic SSOs were engineered with light-removable nucleobase-caging groups.
• Photocleavable backbone linkers were incorporated into the SSOs.
• Optochemical control was demonstrated through light-induced OFF → ON and ON → OFF switching.

Main Results:

• Achieved site-specific installation of light-removable caging groups and photocleavable linkers in SSOs.
• Demonstrated optochemical OFF → ON and ON → OFF switching of SSO activity.
• Enabled precise control over alternative splicing using light in mammalian cells and animals.

Conclusions:

• Light-inducible SSOs provide a powerful tool for optochemical control of alternative splicing.
• This technology offers unprecedented spatial and temporal resolution for gene regulation studies.
• The developed method has potential applications in both cellular research and in vivo studies.