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Phenotype-driven approach to understanding the function of craniofacial regulators using IMPC-generated mouse strains

使用 IMPC 产生的小鼠品系的表型驱动方法来了解颅面调节器的功能

基本信息

批准号：
10806271
负责人：
Jeffrey Ohmann Bush
金额：
$ 2.59万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10806271
关键词：
3-Dimensional Ablation Adult Affect Algorithms Alleles Applications Grants Area Binding Biological Process Biomedical Research Birth Branchial arch structure Candidate Disease Gene Catalogs Categories Cell Adhesion Cell physiology Cells ChIP-seq Classification Cleft Palate Code Congenital Abnormality Craniofacial Abnormalities Cytoskeleton Data Data Set Databases Defect Development E1A-associated p300 protein Ectoderm Effectiveness Embryo Embryonic Development Enhancers Exhibits Experimental Models Face FaceBase Fetus Genes Genetic Genome Goals Health Homeostasis House mice Human Informatics International Knock-out Knockout Mice Knowledge Laboratory mice Life Light Manuals Maxilla Mesenchyme Modeling Molecular Morphogenesis Morphology Mouse Strains Mus Mutant Strains Mice Mutate Mutation Neoplastic Cell Transformation Organism Pattern Perinatal mortality demographics Phenotype Physiologic Ossification Pregnancy Premaxillary palate Process Proteins Reporting Research Role Secondary Palate Shapes Signaling Molecule Structural Congenital Anomalies Syndrome Time Tissues Transgenic Organisms United States National Institutes of Health Validation conditional mutant craniofacial craniofacial complex craniofacial development cranium design experience genome-wide malformation mammalian genome microCT model organism mouse genome mutant phenome phenotypic data response suture fusion transcription factor transcriptome transcriptome sequencing

项目摘要

ABSTRACT Many of the datasets resulting from genome-wide approaches lack functional validation in living organisms. While the laboratory mouse is often used as an experimental model, a large number of mouse genes have unknown functions. The International Mouse Phenotyping Consortium (IMPC) is building the first catalogue of mammalian genome function by generating knockout (KO) mouse strains for every protein-coding gene in the genome. Taking advantage of this opportunity, the two co-PIs have designed this proposal in response to NIH PAR-17-005 for phenotyping IMPC embryonic and perinatal lethal KO mouse lines. Our focus is on mutations that affect the craniofacial complex, based on our expertise in modeling craniofacial malformations in the mouse. While craniofacial defects represent one third of all human birth defects, our knowledge of the underlying cellular and molecular mechanisms remains poor. To select mutant mouse lines for in-depth phenotyping of craniofacial abnormalities, we generated algorithms to intersect the current list of IMPC lethal /subviable KO strains exhibiting craniofacial defects with: 1) all genes present in transcriptomes of mouse embryonic craniofacial domains that were either generated in our labs or available in the FaceBase database; and 2) all significant ChIP-Seq peaks for binding of the enhancer-associated protein p300 in mouse embryonic whole faces present in the FaceBase database. By searching the Mouse Genome Informatics (MGI) database for phenotypic data and by prioritizing genes that have unknown or poorly defined roles in craniofacial develop- ment, we restricted the number of chosen genes to N=30. These comprise regulators of various cellular functions that cause embryonic lethality and craniofacial defects when disrupted in the mouse. Preliminary phenotyping of the KO mouse line for Zfhx4, a gene that exhibited highest maxillary enrichment in our RNA- Seq dataset, revealed that all Zfhx4 KO embryos present cleft palate and maxillary hypoplasia, providing proof of concept for the effectiveness of the proposed strategy. Accordingly, we will characterize 30 mutant lines (15 per lab over 5 years) via the following specific aims: AIM 1: Examination and classification of 30 IMPC mutant mouse lines for craniofacial phenotypes. Employing a two-stage phenotyping pipeline, we will categorize the craniofacial defects for each line and narrow the time of onset. AIM 2. Deep characterization of craniofacial phenotypes. Based on three phenotyping platforms, we will uncover the function(s) of the 30 chosen genes in craniofacial development through in-depth analyses. Platform A will characterize early craniofacial anomalies including defects in branchial arch patterning, as well as primary palate morphogenesis and fusion (E8.5-11.5); Platform B will characterize later abnormalities of secondary palate fusion (E11.5-15.5); and Platform C will dissect perturbations of craniofacial shape/morphology and skull ossification. Shedding light on the cellular and molecular processes controlled by genes that are essential for embryonic development and cause human birth defects when disrupted will be vital to the health of the fetus before and long after birth.

摘要