GENETIC CHARACTERIZATION OF A NOVEL MODEL OF CLEFT PALATE
Stephen A Murray
Jackson Laboratory, 600 Main St, Bar Harbor, Me 04609-1500
Grant 5R03DE019451-02 from National Institute Of Dental & Craniofacial Research
Abstract: Orofacial clefting is one of the most common birth defects in humans, affecting approximately 1 in 700 live births. This frequency likely stems from the complexity of craniofacial morphogenesis, which requires precise regulation of gene expression changes, alterations in cell physiology and morphogenic movements. Although an increasing number of genes have been linked to cleft lip and cleft palate, the mechanisms governing orofacial malformations remain unclear. The overall objective of this project is to expand our understanding of the genetic basis of orofacial clefting through the use of a novel mouse model of cleft palate. The ENU-induced clfp4 mouse harbors a recessive mutation causing cleft secondary palate, omphalocele and skeletal malformations with high penetrance. While simultaneous presentation of orofacial clefting and body wall defects is observed in human syndromes, there are few mouse models that recapitulate these phenotypes. Preliminary mapping data places the mutation at the distal end of Chromosome 10, a region where this combination of phenotypes has not been reported. Thus, clfp4 likely represents a new model of craniofacial/body wall birth defects. Although preliminary phenotypic analysis has revealed that cleft palate in this mutant results from a failure of palate shelf elevation, efficient design and interpretation of more detailed experiments, such as marker analysis, would be greatly facilitated by identification of the clfp4 causative gene. To develop this unique model of cleft palate and to elucidate the mechanism underlying the phenotype, we propose to identify the gene that underlies the cflp4 mutation through high-resolution genetic mapping and targeted re-sequencing, and perform basic phenotypic analysis to begin to understand the mechanistic basis of the defect. This work will improve our understanding of the molecular networks that regulate palatogenesis, providing a foundation for the development of new methods for the diagnosis, prevention and treatment of cleft palate and other craniofacial disorders. Orofacial clefting is one of the most common birth defects in humans, affecting 1 in 700 live births. The total health care costs for surgical repair and post-operative therapies is staggering, and the disfigurement of such disorders often have devastating social and psychological consequences for the affected individual. This proposal will expand our understanding of the etiology of orofacial clefting using a novel mouse model of cleft palate and craniofacial malformations, providing a foundation for the development of new methods for the diagnosis, prevention and treatment of cleft palate and other craniofacial disorders
Keywords: Affect; Apoptosis; Apoptosis Pathway; Birth Defects; Candidate Disease Gene; Candidate Gene; Causality; Cell Death, Programmed; Cell Function; Cell Process; Cell physiology; Cellular Function; Cellular Physiology; Cellular Process; Chromosome 10; Chromosome Mapping; Chromosomes, Human, Pair 10; Cleaved cell; Cleft Lip; Cleft Palate; Congenital Abnormality; Congenital Anatomic Abnormality; Congenital Anatomical Abnormality; Congenital Defects; Congenital Deformity; Congenital Malformation; Congenital omphalocele; Culturing, in vitro Organ; Culturing, in vitro Vertebrate, Organ; Data; Defect; Development; Diagnosis; Disease; Disorder; Distal; Embryo; Embryonic; Etiology; FLR; Failure (biologic function); Foundations; Frequencies (time pattern); Frequency; Future; Gene Action Regulation; Gene Expression; Gene Expression Regulation; Gene Localization; Gene Mapping; Gene Mapping, Total Human and Non-Human; Gene Regulation; Gene Regulation Process; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genetics, Gene Mapping; Goals; Harelip; Health Care Costs; Health Costs; Healthcare Costs; Human; Human, General; Individual; Lesion; Link; Linkage Mapping; Live Birth; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Methods; Mice; Modeling; Molecular; Molecular Genetic Abnormality; Morphogenesis; Movement; Murine; Mus; Mutation; Omphaloceles; Operation; Operative Procedures; Operative Surgical Procedures; Organ Culture; Organ Culture Techniques; Palate; Penetrance; Phenotype; Post-Operative; Postoperative; Postoperative Period; Prevention; Public Health; Reporting; Research Proposals; Resolution; Secondary Palate; Staging; Subcellular Process; Surgical; Surgical Interventions; Surgical Procedure; Syndrome; Time; Work; base; body movement; cleaved; craniofacial; craniofacies; design; designing; disease causation; disease etiology; disease/disorder; disease/disorder etiology; disorder etiology; exome; experiment; experimental research; experimental study; failure; genetic mapping; genome mutation; hare lip; improved; insight; malformation; mouse model; mutant; novel; orofacial; palatogenesis; psychologic; psychological; public health medicine (field); public health relevance; repair; repaired; research study; skeletal; social; stem; surgery
Relevance: RELEVANCE TO PUBLIC HEALTH Orofacial clefting is one of the most common birth defects in humans, affecting 1 in 700 live births. The total health care costs for surgical repair and post-operative therapies is staggering, and the disfigurement of such disorders often have devastating social and psychological consequences for the affected individual. This proposal will expand our understanding of the etiology of orofacial clefting using a novel mouse model of cleft palate and craniofacial malformations, providing a foundation for the development of new methods for the diagnosis, prevention and treatment of cleft palate and other craniofacial disorders
Project start date: 2009-09-01
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PAR-07-418
5R03DE019451-02 (2010): $129195
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Stephen A Murray
GENETIC CHARACTERIZATION OF A NOVEL MODEL OF CLEFT PALATE
Stephen A Murray, Associate Research Scientist
Jackson Laboratory, 600 Main St, Bar Harbor, Me 04609-1500
Grant 1R03DE019451-01A1 from National Institute Of Dental & Craniofacial Research
Abstract: Orofacial clefting is one of the most common birth defects in humans, affecting approximately 1 in 700 live births. This frequency likely stems from the complexity of craniofacial morphogenesis, which requires precise regulation of gene expression changes, alterations in cell physiology and morphogenic movements. Although an increasing number of genes have been linked to cleft lip and cleft palate, the mechanisms governing orofacial malformations remain unclear. The overall objective of this project is to expand our understanding of the genetic basis of orofacial clefting through the use of a novel mouse model of cleft palate. The ENU-induced clfp4 mouse harbors a recessive mutation causing cleft secondary palate, omphalocele and skeletal malformations with high penetrance. While simultaneous presentation of orofacial clefting and body wall defects is observed in human syndromes, there are few mouse models that recapitulate these phenotypes. Preliminary mapping data places the mutation at the distal end of Chromosome 10, a region where this combination of phenotypes has not been reported. Thus, clfp4 likely represents a new model of craniofacial/body wall birth defects. Although preliminary phenotypic analysis has revealed that cleft palate in this mutant results from a failure of palate shelf elevation, efficient design and interpretation of more detailed experiments, such as marker analysis, would be greatly facilitated by identification of the clfp4 causative gene. To develop this unique model of cleft palate and to elucidate the mechanism underlying the phenotype, we propose to identify the gene that underlies the cflp4 mutation through high-resolution genetic mapping and targeted re-sequencing, and perform basic phenotypic analysis to begin to understand the mechanistic basis of the defect. This work will improve our understanding of the molecular networks that regulate palatogenesis, providing a foundation for the development of new methods for the diagnosis, prevention and treatment of cleft palate and other craniofacial disorders. Orofacial clefting is one of the most common birth defects in humans, affecting 1 in 700 live births. The total health care costs for surgical repair and post-operative therapies is staggering, and the disfigurement of such disorders often have devastating social and psychological consequences for the affected individual. This proposal will expand our understanding of the etiology of orofacial clefting using a novel mouse model of cleft palate and craniofacial malformations, providing a foundation for the development of new methods for the diagnosis, prevention and treatment of cleft palate and other craniofacial disorders
Keywords: Affect; Apoptosis; Apoptosis Pathway; Birth Defects; Candidate Disease Gene; Candidate Gene; Causality; Cell Death, Programmed; Cell Function; Cell Process; Cell physiology; Cellular Function; Cellular Physiology; Cellular Process; Chromosome 10; Chromosome Mapping; Chromosomes, Human, Pair 10; Cleaved cell; Cleft Lip; Cleft Palate; Congenital Abnormality; Congenital Anatomic Abnormality; Congenital Anatomical Abnormality; Congenital Defects; Congenital Deformity; Congenital Malformation; Congenital omphalocele; Culturing, in vitro Organ; Culturing, in vitro Vertebrate, Organ; Data; Defect; Development; Diagnosis; Disease; Disorder; Distal; Embryo; Embryonic; Etiology; FLR; Failure (biologic function); Foundations; Frequencies (time pattern); Frequency; Future; Gene Action Regulation; Gene Expression; Gene Expression Regulation; Gene Localization; Gene Mapping; Gene Mapping, Total Human and Non-Human; Gene Regulation; Gene Regulation Process; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genetics, Gene Mapping; Goals; Harelip; Health Care Costs; Health Costs; Healthcare Costs; Human; Human, General; Individual; Lesion; Link; Linkage Mapping; Live Birth; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Methods; Mice; Modeling; Molecular; Molecular Genetic Abnormality; Morphogenesis; Movement; Murine; Mus; Mutation; Omphaloceles; Operation; Operative Procedures; Operative Surgical Procedures; Organ Culture; Organ Culture Techniques; Palate; Penetrance; Phenotype; Post-Operative; Postoperative; Postoperative Period; Prevention; Public Health; Reporting; Research Proposals; Resolution; Secondary Palate; Staging; Subcellular Process; Surgical; Surgical Interventions; Surgical Procedure; Syndrome; Time; Work; base; body movement; cleaved; craniofacial; craniofacies; design; designing; disease causation; disease etiology; disease/disorder; disease/disorder etiology; disorder etiology; exome; experiment; experimental research; experimental study; failure; genetic mapping; genome mutation; hare lip; improved; insight; malformation; mouse model; mutant; novel; orofacial; palatogenesis; psychologic; psychological; public health medicine (field); public health relevance; repair; repaired; research study; skeletal; social; stem; surgery
Relevance: RELEVANCE TO PUBLIC HEALTH Orofacial clefting is one of the most common birth defects in humans, affecting 1 in 700 live births. The total health care costs for surgical repair and post-operative therapies is staggering, and the disfigurement of such disorders often have devastating social and psychological consequences for the affected individual. This proposal will expand our understanding of the etiology of orofacial clefting using a novel mouse model of cleft palate and craniofacial malformations, providing a foundation for the development of new methods for the diagnosis, prevention and treatment of cleft palate and other craniofacial disorders
Project start date: 2009-09-01
Project end date: 2011-08-31
Budget start date: 1-SEP-2009
Budget end date: 31-AUG-2010
PFA/PA: PAR-07-418
1R03DE019451-01A1 (2009): $130500
ENHANCING THE UTILITY OF CRE DRIVER LINES
Stephen A Murray
Jackson Laboratory, 600 Main St, Bar Harbor, Me 04609-1500
Grant 5R21RR026117-02 from National Center For Research Resources
Abstract: Large-scale mouse gene targeting projects, such as KOMP, EUCOMM and NorCOMM, promise to deliver a vast number of conditional loxP-flanked alleles to the scientific community. Capitalizing on this resource will require that a large, diverse set of well-characterized Cre driver lines be made available to researchers around the world. The Jackson Laboratory (JAX) houses and distributes the single largest collection of Cre driver strains, including 69 that are currently distributed as live colonies. Despite the best efforts of those developing new Cre lines, the fidelity of Cre activity is not always ideal. Many difficulties have been reported in various Cre lines, including mosaic or incomplete deletion in a target tissue/cell type, inconsistent activity, expression in non-target tissues, and/or Cre-related toxicity. In many cases, however, these data are not reported or available to the potential user. The overall objective of this proposal is to improve the utility of the Cre driver lines by developing a high-throughput pipeline to systematically address these issues, characterize a subset of Cre lines currently housed in the JAX Repository and to provide the information to the scientific community. To accomplish this task, we will develop assays and protocols to extend the characterization of Cre driver line functionality. This will include evaluation of excision in both target and non-target tissues, and also will address toxicity, sex-specific functional transmission, and consistency of excision. Using these protocols, we will then comprehensively characterize a set of Cre driver lines currently available as live colonies in the Jax Repository, which will serve as a model for characterization of all Cre lines in the future. These data are intended to fill in where published information leaves off, providing essential quality control data and facilitating the identification of appropriate lines for potential users. This effort will be integrated with ongoing efforts of Mouse Genome Informatics to house and display Cre line data and by the development of a JAXCre website to disseminate data on Cre driver lines. As the Repository holding the greatest number of Cre driver lines by far, The Jackson Laboratory is in a perfect position to acquire and disseminate this essential information. By developing a high-throughput flow scheme for extended characterization of Cre lines, we will be able to attack this problem, and enhance the value of these lines for the scientific community. The mouse is an invaluable tool for modeling human disease. The tremendous power of the mouse is due in part to our ability to manipulate its genome in a precise manner, and our toolbox to perform these tasks is rapidly expanding. As part of The Jackson Laboratory Repository, this proposal seeks to leverage our mouse and informatic resources to enhance the value of mouse tool strains for the scientific community. Because of our position as a central resource utilized by thousands of scientists around the world, this project promises to have an important impact on the development of new models of human disease
Keywords: Abscission; Address; Alleles; Allelomorphs; Assay; Behavior; Bioassay; Biologic Assays; Biological Assay; Body Tissues; Cells; Collection; Communities; Complement; Complement Proteins; Crossmatching, Tissue; Data; Development; Evaluation; Excision; Exhibits; Extirpation; Future; Gene Targeting; Genome; Goals; Histocompatibility Testing; Housing; Informatics; Investigators; Laboratories; Left; Life; Mammals, Mice; Mediating; Methods; Mice; Modeling; Murine; Mus; Pattern; Position; Positioning Attribute; Protocol; Protocols documentation; Public Health; Publishing; Quality Control; Removal; Reporter; Reporting; Reproducibility; Research Personnel; Research Resources; Researchers; Resource Informatics; Resources; Scheme; Scientist; Staining method; Stainings; Stains; Surgical Removal; Targetings, Gene; Time; Tissue Crossmatchings; Tissue Typing; Tissues; Toxic effect; Toxicities; Transmission; cell type; design; designing; histocompatibility typing; human disease; improved; mouse genome; public health medicine (field); public health relevance; repository; resection; sex; tool; transmission process; web site
Relevance: Relevance to Public Health The mouse is an invaluable tool for modeling human disease. The tremendous power of the mouse is due in part to our ability to manipulate its genome in a precise manner, and our toolbox to perform these tasks is rapidly expanding. As part of The Jackson Laboratory Repository, this proposal seeks to leverage our mouse and informatic resources to enhance the value of mouse tool strains for the scientific community. Because of our position as a central resource utilized by thousands of scientists around the world, this project promises to have an important impact on the development of new models of human disease
Project start date: 2009-09-07
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: PA-07-336
5R21RR026117-02 (2010): $269550
1R21RR026117-01 (2009): $224625