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JoVE Journal
Biochemistry

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A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
 

A Reporter Based Cellular Assay for Monitoring Splicing Efficiency

Article DOI: 10.3791/63014-v
September 15th, 2021

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Summary September 15th, 2021

This protocol describes a minigene reporter assay to monitor the impact of 5´-splice site mutations on splicing and develops suppressor U1 snRNA for the rescue of mutation-induced splicing inhibition. The reporter and suppressor U1 snRNA constructs are expressed in HeLa cells, and splicing is analyzed by primer extension or RT-PCR.

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