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Understanding the role of PRDM1 in limb development and split hand/foot malformation.

了解 PRDM1 在肢体发育和手/足裂畸形中的作用。

基本信息

批准号：
10474619
负责人：
Brittany Truong
金额：
$ 3.14万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10474619
关键词：
Affect Apical Ectodermal Ridge Binding Biological CRISPR/Cas technology Cells Chromatin Chromatin Remodeling Factor Complex Congenital Abnormality DNA DNA Binding DNA Binding Domain Data Defect Development Developmental Biology Digit structure Disease Disease Outcome Ectoderm Embryo Epigenetic Process Ethylnitrosourea Etiology Event Family Fibroblast Growth Factor Future Gene Activation Gene Expression Gene Expression Profile Genes Genetic Genetic Predisposition to Disease Genetic Transcription Goals Growth Hand Human Laboratories Lead Limb Development Limb structure Mesoderm Mus Mutagenesis Mutate Mutation Newborn Infant Orthologous Gene PRDM1 gene Pathogenesis Pathogenicity Pathway interactions Patients Pectoral Phenotype Play Polydactyly Population Positioning Attribute Process Regulatory Element Repression Role SET Domain Severities Signal Transduction Testing TimeLine Tissues Tretinoin Variant Work Zebrafish Zinc Fingers base causal variant cell motility conditional knockout disease phenotype exome sequencing experimental study foot gene regulatory network improved in vivo insight knock-down malformation morphogens mutant negative affect novel null mutation recruit repaired transcription factor transcriptome sequencing variant of unknown significance

项目摘要

PROJECT SUMMARY/ABSTRACT Limb development is strictly regulated by gene regulatory networks governed by a complex network of regulatory elements, transcription factors, and epigenetic modifiers. Each component is interconnected, and misregulation at any point in the developmental timeline can lead to congenital limb defects, which affect 1 in 2,000 newborns. One factor that regulates early limb initiation and outgrowth in both mice and zebrafish is PRDM1, though the mechanism is unknown. Uncovering this has become increasingly more important as we have recently identified three human families with split hand/foot malformation (SHFM) of unknown genetic etiology but potentially pathogenic PRDM1 variants. PRDM1 is a versatile transcription factor, capable of both activation and repression by binding directly to DNA at its zinc finger domain or by recruiting chromatin modifiers to its SET domain to regulate transcription . The DNA-binding domain of PRDM1 is either absent or mutated in zebrafish mutants and in SHFM families. Therefore, I hypothesize that direct DNA binding through the zinc finger domain of PRDM1 is necessary for activation of gene expression required for maintaining AER activity during limb development. The proposed study will test this hypothesis in three focused aims. The first aim will use a conditional rescue experiment to determine the functionally active domain of Prdm1a (zebrafish ortholog) during limb development. Constructs of modified prdm1a, in which different domains are deleted, will be injected into null zebrafish mutants. The second aim will determine where Prdm1a binds to DNA during pectoral fin bud development. The third aim will test the pathogenicity of each SHFM PRDM1 variant in zebrafish following mutagenesis by CRISPR-Cas9 and homology directed repair. I will analyze pectoral fin bud development and DNA binding in each mutant. Together, the results from this study will improve our understanding of one of the many pathways involved in limb development. It will also provide insight into how mutations in PRDM1 can lead to congenital limb disorders and help us better predict disease outcomes or phenotype severity based on the given variant.

项目总结/文摘