Neural crest-specific deletion of Rbfox2 in mice leads to craniofacial abnormalities including cleft palate.
Alternative splicing (AS) creates proteomic diversity from a limited size genome by generating numerous transcripts from a single protein-coding gene. Tissue-specific regulators of AS are essential components of the gene regulatory network, required for normal cellular function, tissue patterning, and embryonic development. However, their cell-autonomous function in neural crest development has not been explored. Here, we demonstrate that splicing factor Rbfox2 is expressed in the neural crest cells (NCCs), and deletion of Rbfox2 in NCCs leads to cleft palate and defects in craniofacial bone development. RNA-Seq analysis revealed that Rbfox2 regulates splicing and expression of numerous genes essential for neural crest/craniofacial development. We demonstrate that Rbfox2-TGF-β-Tak1 signaling axis is deregulated by Rbfox2 deletion. Furthermore, restoration of TGF-β signaling by Tak1 overexpression can rescue the proliferation defect seen in Rbfox2 mutants. We also identified a positive feedback loop in which TGF-β signaling promotes expression of Rbfox2 in NCCs.
Pubmed ID: 31241461 RIS Download
Research resources used in this publication
- Cuffdiff (RRID:SCR_001647) (software resource)
- B6.129S2-Rbfox2tm1.1Dblk/J (RRID:IMSR_JAX:014090) (organism)
- pcDNA3 TAK1/F (RRID:Addgene_44161) (plasmid)
- pCMV5B-Smad4 (RRID:Addgene_11743) (plasmid)
- pCMV5B-HA-Smad2 (RRID:Addgene_11734) (plasmid)
- pCMV5B-Flag-Smad3 (RRID:Addgene_11742) (plasmid)
- 129S4.Cg-E2f1Tg(Wnt1-cre)2Sor/J (RRID:IMSR_JAX:022137) (organism)
- B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato.-EGFP)Luo/J (RRID:IMSR_JAX:007676) (organism)
- B6;129-Pax3tm1(cre)Joe/J (RRID:IMSR_JAX:005549) (organism)
- Cuffdiff (RRID:SCR_001647) (software resource)
- Monoclonal Anti-Actin, α-Smooth Muscle (RRID:AB_476701) (antibody)
Additional research tools detected in this publication
Antibodies used in this publication
- Monoclonal Anti-Actin, α-Smooth Muscle (RRID:AB_476701)
- neurofilament (NF-M) antibody - Jessell, T.M. / Dodd, J.; HHMI/Columbia University (RRID:AB_531793)
- Recombinant Anti-Ki67 antibody [SP6] (RRID:AB_302459)
- Fox2 / RBM9 antibody (RRID:AB_2285090)
- Anti-p38 MAPK Antibody, Unconjugated (RRID:AB_330713)
- Rabbit Anti-p38 MAPK, phospho (Thr180 / Tyr182) Monoclonal Antibody, Unconjugated, Clone 12F8 (RRID:AB_331765)
- Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (RRID:AB_490941)
- Anti-β-Actin Antibody (C4) (RRID:AB_626632)
- Phospho-TAK1 (Ser412) Antibody (RRID:AB_2140096)
- Fibronectin (EP5) (RRID:AB_627598)
- Smad2 (D43B4) XP Rabbit mAb (RRID:AB_10626777)
- Mouse Anti-Tak1 Monoclonal Antibody, Unconjugated, Clone H-5 (RRID:AB_2140220)
Associated grants
- Agency: National Research Foundation Singapore, International
Id: NRF-NRFF2016-01
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This is a list of tools and resources that we have found mentioned in this publication.
Addgene (tool)
RRID:SCR_002037
Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
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Probabilistic framework that quantitates the expression level of alternatively spliced genes from RNA-Seq and identifies differentially regulated isoforms or exons across samples.
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A commercial antibody supplier which supplies primary and secondary antibodies, biochemicals, proteins, peptides, lysates, immunoassays and other kits.
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RRID:AB_476701
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RRID:AB_531793
This monoclonal targets neurofilament (NF-M)
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RRID:AB_302459
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This monoclonal targets RBM9
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RRID:AB_330713
This polyclonal targets p38 MAPK
View all literature mentionsRabbit Anti-p38 MAPK, phospho (Thr180 / Tyr182) Monoclonal Antibody, Unconjugated, Clone 12F8 (antibody)
RRID:AB_331765
This monoclonal targets p38 MAPK, phospho (Thr180 / Tyr182)
View all literature mentionsPhospho-Smad2 (Ser465/467) (138D4) Rabbit mAb (antibody)
RRID:AB_490941
This monoclonal targets Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb
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RRID:AB_626632
This monoclonal targets β-Actin
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RRID:AB_2140096
This polyclonal targets MAP3K7
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RRID:AB_627598
This monoclonal targets FN1
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RRID:AB_10626777
This monoclonal targets Smad2 (D43B4) XP Rabbit mAb
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RRID:SCR_001647
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RRID:IMSR_JAX:014090
Mus musculus with name B6.129S2-Rbfox2tm1.1Dblk/J from IMSR.
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RRID:IMSR_JAX:022137
Mus musculus with name 129S4.Cg-E2f1Tg(Wnt1-cre)2Sor/J from IMSR.
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RRID:IMSR_JAX:007676
Mus musculus with name B6.129(Cg)-Gt(ROSA)26Sortm4(ACTB-tdTomato.-EGFP)Luo/J from IMSR.
View all literature mentionsB6;129-Pax3tm1(cre)Joe/J (organism)
RRID:IMSR_JAX:005549
Mus musculus with name B6;129-Pax3tm1(cre)Joe/J from IMSR.
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RRID:SCR_001647
Software that estimates expression at transcript-level resolution and controls for variability evident across replicate libraries.
View all literature mentionsMonoclonal Anti-Actin, α-Smooth Muscle (antibody)
RRID:AB_476701
This monoclonal targets Actin, α-Smooth Muscle
View all literature mentions
