Melania Barile1,2, 1,2, Isabella Inzani3, Shila Ghazanfar4, Jennifer Nichols2,5, John C. Marioni4,6,7, Carolina Guibentif1,2,8 & Berthold Göttgens ORCID: orcid.org/0000-0001-6302-57051,2 Genome Biology volume 22, Article number: 197 (2021) Cite this article Abstract Background Single-cell technologies are transforming biomedical research, including the recent demonstration that unspliced pre-mRNA present in single-cell RNA-Seq permits prediction of future expression states.