A bipartite boundary element restrictsUBE3Aimprinting to mature neurons

Abstract

Angelman syndrome (AS) is a severe neurodevelopmental disorder caused by the loss of function from the maternal allele ofUBE3A, a gene encoding an E3 ubiquitin ligase.UBE3Ais only expressed from the maternally inherited allele in mature human neurons due to tissue-specific genomic imprinting. Imprinted expression ofUBE3Ais restricted to neurons by expression ofUBE3A antisense transcript(UBE3A-ATS) from the paternally inherited allele, which silences the paternal allele ofUBE3Aincis. However, the mechanism restrictingUBE3A-ATSexpression andUBE3Aimprinting to neurons is not understood. We used CRISPR/Cas9-mediated genome editing to functionally define a bipartite boundary element critical for neuron-specific expression ofUBE3A-ATSin humans. Removal of this element led to up-regulation ofUBE3A-ATSwithout repressing paternalUBE3A. However, increasing expression ofUBE3A-ATSin the absence of the boundary element resulted in full repression of paternalUBE3A, demonstrating thatUBE3Aimprinting requires both the loss of function from the boundary element as well as the up-regulation ofUBE3A-ATS. These results suggest that manipulation of the competition betweenUBE3A-ATSandUBE3Amay provide a potential therapeutic approach for AS.