Comparison of TCF4 repeat expansion length in corneal endothelium and leukocytes of patients with Fuchs endothelial corneal dystrophy
| Autor: | Shubham Basu, Keith H. Baratz, Leo J. Maguire, Sanjay V. Patel, Ross A. Aleff, Michael P. Fautsch, Eric D. Wieben, Tommy A. Rinkoski |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Male
Eye Diseases Molecular biology Polymerase Chain Reaction Electrophoretic Blotting Cornea chemistry.chemical_compound White Blood Cells Medical Conditions Sequencing techniques Transcription Factor 4 Animal Cells Medicine and Health Sciences Leukocytes Child Gel Electrophoresis Aged 80 and over Multidisciplinary Endothelium Corneal RNA sequencing TCF4 Middle Aged Blotting Southern Corneal Disorders Medicine Female Anatomy Cellular Types Research Article Corneal endothelium Genotype Science Ocular Anatomy Immune Cells Immunology Molecular Probe Techniques Surgical and Invasive Medical Procedures Biology Biomolecular isolation Electrophoretic Techniques Ocular System Humans Genetic Predisposition to Disease Endothelium Gene Transcription factor Southern blot Aged Blood Cells Fuchs' Endothelial Dystrophy Biology and Life Sciences Cell Biology DNA DNA isolation eye diseases Research and analysis methods Ophthalmology Molecular biology techniques chemistry Cardiovascular Anatomy sense organs Trinucleotide repeat expansion Trinucleotide Repeat Expansion Southern Blot Fuchs Endothelial Corneal Dystrophy |
| Zdroj: | PLoS ONE PLoS ONE, Vol 16, Iss 12, p e0260837 (2021) |
| ISSN: | 1932-6203 |
| Popis: | Expansion of CTG trinucleotide repeats (TNR) in the transcription factor 4 (TCF4) gene is highly associated with Fuchs Endothelial Corneal Dystrophy (FECD). Due to limitations in the availability of DNA from diseased corneal endothelium, sizing of CTG repeats in FECD patients has typically been determined using DNA samples isolated from peripheral blood leukocytes. However, it is non-feasible to extract enough DNA from surgically isolated FECD corneal endothelial tissue to determine repeat length based on current technology. To circumvent this issue, total RNA was isolated from FECD corneal endothelium and sequenced using long-read sequencing. Southern blotting of DNA samples isolated from primary cultures of corneal endothelium from these same affected individuals was also assessed. Both long read sequencing and Southern blot analysis showed significantly longer CTG TNR expansion (>1000 repeats) in the corneal endothelium from FECD patients than those characterized in leukocytes from the same individuals (TCF4 CTG repeat expansions in the FECD corneal endothelium are much longer than those found in leukocytes. |
| Databáze: | OpenAIRE |
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