Data CitationsLukacs M, Chatuverdi P, Stottmann R. and mutant (H) RNA in situ hybridization with -feeling (mutant (J) RNA in situ hybridization with -sense (having a hypo-morphic mutation in mutation decreases surface GPI manifestation. Surprisingly, showed tissue-specific manifestation with enrichment in the brain and face. We found the phenotype is due to apoptosis of neural crest cells (NCCs) and the cranial neuroepithelium. We showed folinic acid supplementation can partially save the cleft lip phenotype. Finally, we generated a novel mouse model of NCC-specific total GPI deficiency. These mutants developed median cleft lip and palate demonstrating a previously Rabbit Polyclonal to MCM3 (phospho-Thr722) undocumented cell autonomous part for GPI biosynthesis in NCC development. lack stable surface Syncytial Virus Inhibitor-1 expression of a variety of GPI-anchored proteins (GPI-APs) (Kinoshita, 2014; Hansen Syncytial Virus Inhibitor-1 et al., 2013). Autosomal recessive mutations in cause Hyperphosphatasia with Mental Retardation 3 (HPMRS3 OMIM # 614207), an IGD that presents with variably penetrant hyperphosphatasia, developmental delay, seizures, microcephaly, heart defects, and a variety of neurocristopathies including Hirschsprungs disease, cleft Syncytial Virus Inhibitor-1 lip, cleft palate, and facial dysmorphia (Hansen et al., 2013; Jezela-Stanek et al., 2016; Krawitz et al., 2013; Naseer et al., 2016). Currently, there is no known molecular mechanism to explain the cause of these phenotypes or therapies for these individuals. In a ahead genetic ENU mutagenesis display, we previously recognized the mouse mutant with Cleft Lip, Cleft Palate, Edema, and Exencephaly (To day, embryonic phenotypes of GPI biosynthesis mutants have been difficult to study due to the early lethal phenotypes associated with germline knockout of GPI biosynthesis genes (McKean and Niswander, 2012; Nozaki et al., 1999; Mohun et al., 2013; Zoltewicz et al., 1999). In this study, we took advantage of the hypo-morphic mutant and a conditional knockout of GPI biosynthesis to determine the mechanism of the various phenotypes and tested the hypothesis that GPI-anchored Folate Receptor 1 (FOLR1) is responsible for the phenotypes observed. Results The mutant phenotype is definitely caused by a missense mutation in mutant inside a mouse N-ethyl-N-nitrosourea (ENU) mutagenesis display for recessive alleles leading to organogenesis phenotypes (Stottmann et al., 2011). homozygous mutants displayed multiple partially penetrant phenotypes. Inside a subset of 70 mutants from late organogenesis phases (~E16.5-E18.5), we noted cranial neural tube problems (exencephaly) in 61 (87%), cleft lip in 22 (31%), cleft palate in 13 (19%), and edema in six embryos (9%) (Number 1ACH). Skeletal preparations of mutants recognized a defect in frontal bone ossification (Number 1ICL, n?=?5/5 mutants) and a statistically significant decrease in limb size (Number 1MCP). We previously reported a genetic mapping strategy with the Mouse Common Genotyping Array which recognized a 44 Mb region of homozygosity for the mutagenized A/J genome on chromosome 7 (Number 1Q) (Stottmann et al., 2011). We then took a whole exome sequencing approach and sequenced 3 homozygous mutants. Analysis of single foundation pair variants which were homozygous in all three mutants with expected high effect as determined by the sequence analysis pipeline (e.g. missense variants in conserved residue, premature quit codons, etc.) and not already known strain polymorphisms in dbSNP remaining only one candidate variant Syncytial Virus Inhibitor-1 (Table 1). This was a homozygous missense mutation in the initiating methionine (c.A1G, p.M1V) in exon.