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Iranian Journal of Biotechnology، جلد ۲۱، شماره ۴، صفحات ۴۸-۵۸
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عنوان انگلیسی CRISPR/Cas9-Induced Fam83h Knock-out Leads to Impaired Wnt/ β-Catenin Pathway and Altered Expression of Tooth Mineralization Genes in Mice
چکیده انگلیسی مقاله Background: Dental enamel formation is a complex process that is regulated by various genes. One such gene, Family 
With Sequence Similarity 83 Member H (Fam83h), has been identified as an essential factor for dental enamel formation. 
Additionally, Fam83h has been found to be potentially linked to the Wnt/β-catenin pathway.
Objectives: This study aimed to investigate the effects of the Fam83h knockout gene on mineralization and formation of 
teeth, along with mediators of the Wnt/β-catenin pathway as a development aspect in mice.
Materials and Methods: To confirm the Fam83h-KnockOut mice, both Sanger sequencing and Western blot methods 
were used. then used qPCR to measure the expression levels of genes related to tooth mineralization and formation 
of dental root, including Fam20a, Dspp, Dmp1, Enam, Ambn, Sppl2a, Mmp20, and Wnt/β-catenin pathway mediators, 
in both the Fam83h-Knockout and wild-type mice at 5, 11 and 18 days of age. also the expression level of Fgf10 and 
mediators of the Wnt/β-catenin pathway was measured in the skin of both Knockout and wild-type mice using qPCR. A 
histological assessment was then performed to further investigate the results.
Results: A significant reduction in the expression levels of Ambn, Mmp20, Dspp, and Fgf10 in the dental root of Fam83h-Knockout 
mice compared to their wild-type counterparts was demonstrated by our results, indicating potential disruptions in tooth development. 
Significant down-regulation of CK1a, CK1e, and β-catenin in the dental root of Fam83h-Knockout mice was associated with a 
reduction in mineralization and formation-related gene. Additionally, the skin analysis of Fam83h-Knockout mice revealed reduced 
levels of Fgf10, CK1a, CK1e, and β-catenin. Further histological assessment confirmed that the concurrent reduction of Fgf10 
expression level and Wnt/β-catenin genes were associated with alterations in hair follicle maturation.
Conclusions: The concurrent reduction in the expression level of both Wnt/β-catenin mediators and mineralization-related 
genes, resulting in the disruption of dental mineralization and formation, was caused by the deficiency of Fam83h. Our 
findings suggest a cumulative effect and multi-factorial interplay between Fam83h, Wnt/Β-Catenin signaling, and dental 
mineralization-related genes subsequently, during the dental formation process. 
کلیدواژه‌های انگلیسی مقاله Amelogenesis Imperfecta,CRISPR-Cas Systems,Family with sequence similarity 83 member H protein,Fgf10 protein,Wnt Signaling Pathway
نویسندگان مقاله NASSERI SHERKO |
Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

Sara Parsa |
Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

Zakaria Vahabzadeh |
Liver and Digestive Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

Babak Baban |
Department of Oral Biology and Diagnostic Sciences, Dental College of Georgia, Augusta University, Augusta, Georgia

Mohammad Bagher Khadem-erfan |
Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

Bahram Nikkhoo |
Department of Pathology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran

Mohammad Rastegar Khosravi |
Department of Endodontics, Faculty of Dentistry, Kurdistan University of Medical Sciences, Sanandaj, Iran

Saman Bahrami |
Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

Fardin Fathi |
Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

نشانی اینترنتی https://www.ijbiotech.com/article_179906_0763fc9c43e297ecbb6426549a47753d.pdf
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