Based on studies in mice, the homeobox transcription factor HOXA2, which directly binds to an evolutionarily conserved site in the second intron of the ROBO2 gene, is needed for the maintenance of ROBO2 expression during pontine neuron migration (Geisen et al. 2008).

Also based on mouse studies, LHX2, a LIM-homeodomain transcription factor, directly represses transcription of the ROBO2 gene by binding to evolutionarily conserved LHX2 binding sites about 50 kb downstream from the ROBO2 gene transcription start site. LHX2 is involved in thalamocortical axon guidance (Marcos-Mondejar et al. 2012). In commissural relay neurons of the dorsal spinal cord, however, ROBO2 expression is not affected by LHX2 (Wilson et al. 2008).

In zebrafish, transcription of Robo2 is directly stimulated by Mecp2 (Leong et al. 2015).