title: RBFOX1 and RBFOX2 are dispensable in iPSCs and iPSC-derived neurons and do not contribute to neural-specific paternal UBE3A silencing.
abstract: Angelman Syndrome (AS) is a rare neurodevelopmental disorder caused by loss of function of the maternally inherited copy of UBE3A, an imprinted gene expressed biallelically in most tissues, but expressed exclusively from the maternal allele in neurons. Active transcription of the neuron-specific long non-coding RNA (lncRNA), UBE3A-ATS, has been shown to silence paternal UBE3A. We hypothesized that alternative splicing factors RBFOX2 and RBFOX1 might mediate splicing changes and result in the transcription of UBE3A-ATS in neurons. We found that RBFOX2 and RBFOX1 both bind to UBE3A-ATS transcript in neurons, but are not required for gene expression and/or neuron-specific processing in the SNURF/SNRPN-UBE3A region. However, we found that depletion of RBFOX2 causes a proliferation phenotype in immature neural cultures, suggesting that RBFOX2 is involved in division versus differentiation decisions in iPSC-derived neural progenitors. Absence of RBFOX2 also altered the expression of some genes that are important for glutamatergic neocortical development and Wnt-Frizzled signalling in mature neuronal cultures. Our data show that while RBFOX1 and RBFOX2 do not mediate neuron-specific processing of UBE3A-ATS, these proteins play important roles in developing neurons and are not completely functionally redundant.
authors.affiliation: Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, Connecticut, 06030, USA.
authors.forename: Pin-Fang
authors.initials: PF
authors.lastname: Chen
authors.affiliation: Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, Connecticut, 06030, USA.
authors.forename: Jack S
authors.initials: JS
authors.lastname: Hsiao
authors.affiliation: Department of Neuroscience, University of Connecticut Health Center, Farmington, Connecticut, 06030, USA.
authors.forename: Carissa L
authors.initials: CL
authors.lastname: Sirois
authors.affiliation: Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, Connecticut, 06030, USA.|University of Connecticut Stem Cell Institute, University of Connecticut Health Center, Farmington, Connecticut, 06030, USA.
authors.forename: Stormy J
authors.initials: SJ
authors.lastname: Chamberlain
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology; D018392:Genomic Imprinting; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D000072260:RNA Splicing Factors / Q000235:genetics / Q000378:metabolism*; D062085:RNA, Long Noncoding / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D060449:Wnt Signaling Pathway
chemical_list: C455479:RBFOX1 protein, human; C454924:RBFOX2 protein, human; D000072260:RNA Splicing Factors; D062085:RNA, Long Noncoding; D012097:Repressor Proteins