- Kemmler, Cassie L;
- Smolikova, Jana;
- Moran, Hannah R;
- Mannion, Brandon J;
- Knapp, Dunja;
- Lim, Fabian;
- Czarkwiani, Anna;
- Aguayo, Viviana Hermosilla;
- Rapp, Vincent;
- Fitch, Olivia E;
- Bötschi, Seraina;
- Selleri, Licia;
- Farley, Emma;
- Braasch, Ingo;
- Yun, Maximina;
- Visel, Axel;
- Osterwalder, Marco;
- Mosimann, Christian;
- Kozmik, Zbynek;
- Burger, Alexa
The cell type-specific expression of key transcription factors is central to development. Brachyury/T/TBXT is a major transcription factor for gastrulation, tailbud patterning, and notochord formation; however, how its expression is controlled in the mammalian notochord has remained elusive. Here, we identify the complement of notochord-specific enhancers in the mammalian Brachyury/T/TBXT gene. Using transgenic assays in zebrafish, axolotl, and mouse, we discover three Brachyury -controlling notochord enhancers T3, C , and I in human, mouse, and marsupial genomes. Acting as Brachyury-responsive, auto-regulatory shadow enhancers, deletion of all three enhancers in mouse abolishes Brachyury/T expression selectively in the notochord, causing specific trunk and neural tube defects without gastrulation or tailbud defects. Sequence and functional conservation of Brachyury -driving notochord enhancers with the brachyury/tbxtb loci from diverse lineages of fishes dates their origin to the last common ancestor of jawed vertebrates. Our data define the enhancers for Brachyury/T/TBXTB notochord expression as ancient mechanism in axis development.