RBFOX1 and RBFOX2 are dispensable in iPSCs and iPSC-derived neurons and do not contribute to neural-specific paternal UBE3A silencing

Chen, Pin Fang; Hsiao, Jack S.; Sirois, Carissa L.; Chamberlain, Stormy J.

doi:10.1038/srep25368

Cited by 20 publications

(12 citation statements)

References 43 publications

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“…All Rbfox proteins recognize the same sequence motif and regulate alternative splicing of an overlapping set of target exons indicating functional redundancy [6][7][8]. Functional differences between Rbfox1, Rbfox2, and Rbfox3 exist as well [9]. In agreement with functional specificity the brain-specific Rbfox1 knockout mouse develops spontaneous epilepsy [10], whereas the brainspecific Rbfox2 knockout mouse shows impairments in cerebellar development and motor function [6].…”

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confidence: 99%

Identification of a classic nuclear localization signal at the N terminus that regulates the subcellular localization of Rbfox2 isoforms during differentiation of NMuMG and P19 cells

et al. 2016

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Edited by Claus AzzalinNuclear localization of the alternative splicing factor Rbfox2 is achieved by a C-terminal nuclear localization signal (NLS) which can be excluded from some Rbfox2 isoforms by alternative splicing. While this predicts nuclear and cytoplasmic localization, Rbfox2 is exclusively nuclear in some cell types. Here, we identify a second NLS in the N terminus of Rbfox2 isoform 1A that is not included in Rbfox2 isoform 1F. Rbfox2 1A isoforms lacking the Cterminal NLS are nuclear, whereas equivalent 1F isoforms are cytoplasmic. A shift in Rbfox2 expression toward cytoplasmic 1F isoforms occurs during epithelial to mesenchymal transition (EMT) and could be important in regulating the activity and function of Rbfox2.

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confidence: 99%

Identification of a classic nuclear localization signal at the N terminus that regulates the subcellular localization of Rbfox2 isoforms during differentiation of NMuMG and P19 cells

et al. 2016

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“…Addition to be as critical determinant for neuronal differentiation or neurogenesis in NSCs or NPCs, lncRNAs are also acting as pivotal regulatory molecules in several neurodevelopmental related diseases including Huntington's disease (HD) [71], Autism spectrum disorder (ASD) [72], Angelman syndrome (AS) [73], vascular disorders induced ischemic stroke [74,75]. LncRNA Tcl1 Upstream Neuron-Associated lincRNA (TUNA) was found to be associated with HD, which function for maintenance of pluripotency and neural differentiation by interaction with three RBPs [71].…”

Section: The Effect Of Lncrna On Neurodevelopmental Disorder Via Targmentioning

confidence: 99%

The functions of long non-coding RNAs in neural stem cell proliferation and differentiation

et al. 2020

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The capacities for neural stem cells (NSCs) self-renewal with differentiation are need to be precisely regulated for ensuring brain development and homeostasis. Recently, increasing number of studies have highlighted that long non-coding RNAs (lncRNAs) are associated with NSC fate determination during brain development stages. LncRNAs are a class of non-coding RNAs more than 200 nucleotides without protein-coding potential and function as novel critical regulators in multiple biological processes. However, the correlation between lncRNAs and NSC fate decision still need to be explored in-depth. In this review, we will summarize the roles and molecular mechanisms of lncR-NAs focusing on NSCs self-renewal, neurogenesis and gliogenesis over the course of neural development, still more, dysregulation of lncRNAs in all stage of neural development have closely relationship with development disorders or glioma. In brief, lncRNAs may be explored as effective modulators in NSCs related neural development and novel biomarkers for diagnosis and prognosis of neurological disorders in the future.

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“…All CRISPRs were designed using MIT's CRISPR design tool (http://crispr.mit.edu). sgRNAs were cloned into the pX459v2.0 vector (Addgene 62988), as previously described 21,22 unless otherwise indicated. The 3' sgRNA used to generate the UBE3A KO line was designed by the hESC/iPSC Targeting Core at the University of Connecticut Health Center and was cloned into the pX330 vector.…”

Section: Crispr/cas9-mediated Genome Editingmentioning

confidence: 99%

Abundance and localization of human UBE3A protein isoforms

Sirois¹,

Bloom²,

Fink

et al. 2020

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Loss of UBE3A expression, a gene regulated by genomic imprinting, causes Angelman Syndrome (AS), a rare neurodevelopmental disorder. The UBE3A gene encodes an E3 ubiquitin ligase with three known protein isoforms in humans. Studies in mouse suggest that the human isoforms may have differences in localization and neuronal function. A recent case study reported mild AS phenotypes in individuals lacking one specific isoform. Here we have used CRISPR/Cas9 to generate isogenic human embryonic stem cells (hESCs) that lack the individual protein isoforms. We demonstrate that isoform 1 accounts for the majority of UBE3A protein in hESCs and neurons. We also show that UBE3A predominantly localizes to the cytoplasm in both wild type and isoform-null cells. Finally, we show that neurons lacking isoform 1 display a less severe electrophysiological AS phenotype.Angelman Syndrome (AS) is a rare neurodevelopmental disorder that affects 1 in 15,000 individuals, and is characterized by severe seizures, intellectual disability, absent speech, ataxia, and happy affect 1 . AS is caused by loss of function from the maternally-inherited copy of UBE3A. UBE3A is regulated by tissue-specific genomic imprintingit is expressed exclusively from the maternal allele in neurons and is expressed biallelically in other cell types 2,3 . UBE3A encodes an E3 ubiquitin ligase that forms polyubiquitin chains to substrates, targeting them for degradation by the 26S proteasome 4 . In humans, there are three known UBE3A protein isoforms 5 , all of which include the Homologous to E6AP Carboxy Terminus (HECT) domain and are thus capable of functioning as an E3 ligase. Human isoforms 2 and 3 only differ from human isoform 1 by 23 and 20 amino acids, respectively, at their N termini 6 (Fig 1a). While there has been little published research examining the human protein isoforms in human cells, human isoforms ectopically expressed in mouse indicate that the isoforms likely have differences in localization and function 7-9 . Furthermore, it is currently unknown whether the different human isoforms have different abundances or functions. This knowledge is important as some therapeutic avenues currently being explored for AS involve delivery and expression of exogenous UBE3A transgenes using vector-based delivery 10 . For these approaches, knowledge of which of the protein isoforms need to be replaced in AS is essential.Recently, Sadhwani and colleagues published three cases of AS in two different families caused by missense mutations at the isoform 1 translational start site 11 . Interestingly, these patients present with milder phenotypes than AS patients with 15q11-q13 deletions or other UBE3A loss of function mutations, including normal gait and use of syntactic speech. This data, coupled with the fact that the human and mouse isoforms are not entirely conserved, illustrates the need to study the human UBE3A protein isoforms specifically, as they may play a role in both normal neuronal function and in disease. Here, we have used human embryonic stem cells (h...

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RBFOX1 and RBFOX2 are dispensable in iPSCs and iPSC-derived neurons and do not contribute to neural-specific paternal UBE3A silencing

Cited by 20 publications

References 43 publications

Identification of a classic nuclear localization signal at the N terminus that regulates the subcellular localization of Rbfox2 isoforms during differentiation of NMuMG and P19 cells

Identification of a classic nuclear localization signal at the N terminus that regulates the subcellular localization of Rbfox2 isoforms during differentiation of NMuMG and P19 cells

The functions of long non-coding RNAs in neural stem cell proliferation and differentiation

Abundance and localization of human UBE3A protein isoforms

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