SUMMARY The abscission checkpoint pauses the progression of cytokinetic abscission at the intercellular bridge when various checkpoint-sustaining errors are sensed. A unique cytoplasmic organelle, the abscission checkpoint body (ACB), forms when the abscission checkpoint is active. This organelle houses proteins involved in abscission and RNA splicing factors. Here, we investigate the role of RNA splicing factors in abscission checkpoint maintenance. A specific subset of RNA splicing factors localizes to ACBs, while others move back into the nucleus. The key RNA splicing factor SF3b1 is aberrantly localized in the cytoplasm and its catalytic activity limited when ACBs are present. Efficient abscission was found to rely on RNA splicing, and we have identified patterns in gene expression and RNA splicing that occur at this time in cell division. Finally, we found a unique RNA landscape associated with the formation of ACBs due to a specific set of changes in RNA expression and alternative splicing, revealing a previously unrecognized level of regulation imparted by abscission checkpoint activity.