Alternative RNA processing and strategies to modulate splicing

Abstract

[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT REQUEST OF AUTHOR.] Spinal Muscular Atrophy (SMA) is a genetic disease that is intrinsically linked to an alternative RNA splicing event that leads a defective protein. To better understand how aberrant RNA processing events occur and lead to disease development, we studied cis and trans elements associated with survival motor neuron (SMN) pre-mRNA splicing. As a means to move more towards a translational approach to restore proper SMN pre-mRNA splicing, we developed a negatively-acting bifunctional RNA that was shown in several different assays to function both in vitro and in the more complex environment of the SMA mouse model in vivo. To further extend our understanding of pre-mRNA processing and the factors involved in its regulation, we utilize the Minute Virus of Mice (MVM) as a model system to study the fundamental process of alternative splicing. We examined the cis elements within the MVM genome that control the expression of an exon by placing the elements in a heterologous context. We have also identified trans elements involved in MVM alternative splicing. Taken together, this research examined the viral life cycle by analyzing the critical steps in alternative splicing involved in a viable MVM infection, and will also give insights into the fundamental biological process of alternative splicing.