Lampe1: An ENU-Germline Mutation Causing Spontaneous Hepatosteatosis Identified through Targeted Exon-Enrichment and Next-Generation Sequencing
| Autor: | Jorge A. Bezerra, Shiva Kumar Shanmukhappa, Mehdi Keddache, Kasper Hoebe, Patrick Putnam, Kristin Lampe, Senad Divanovic, Rachel Sheridan |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Genetic Screens
Mutant Gene Identification and Analysis lcsh:Medicine Nonalcoholic Steatohepatitis medicine.disease_cause Biochemistry Germline Exon Mice 0302 clinical medicine lcsh:Science Sequence Deletion Genetics 0303 health sciences Mutation Multidisciplinary Liver Diseases Liver Neoplasms Exons Phenotype Oncology Liver 030220 oncology & carcinogenesis Gene Targeting Medicine Research Article Sequence analysis Mutagenesis (molecular biology technique) Gastroenterology and Hepatology Biology Molecular Genetics 03 medical and health sciences Germline mutation Genetic Mutation Gastrointestinal Tumors medicine Cancer Genetics Animals Humans Gene Germ-Line Mutation 030304 developmental biology lcsh:R Cancers and Neoplasms Hepatocellular Carcinoma Sequence Analysis DNA Lipid Metabolism Molecular biology Mice Mutant Strains Fatty Liver Mice Inbred C57BL Metabolism Mutagenesis Ethylnitrosourea Genetics of Disease lcsh:Q RNA Splice Sites Gene Function Animal Genetics |
| Zdroj: | PLoS ONE PLoS ONE, Vol 6, Iss 7, p e21979 (2011) |
| ISSN: | 1932-6203 |
| Popis: | Using a small scale ENU mutagenesis approach we identified a recessive germline mutant, designated Lampe1 that exhibited growth retardation and spontaneous hepatosteatosis. Low resolution mapping based on 20 intercrossed Lampe1 mice revealed linkage to a ∼14 Mb interval on the distal site of chromosome 11 containing a total of 285 genes. Exons and 50 bp flanking sequences within the critical region were enriched with sequence capture microarrays and subsequently analyzed by next-generation sequencing. Using this approach 98.1 percent of the targeted DNA was covered with a depth of 10 or more reads per nucleotide and 3 homozygote mutations were identified. Two mutations represented intronic nucleotide changes whereas one mutation affected a splice donor site in intron 11-12 of Palmitoyl Acetyl-coenzyme A oxygenase-1 (Acox1), causing skipping of exon 12. Phenotyping of Acox1(Lampe1) mutants revealed a progression from hepatosteatosis to steatohepatitis, and ultimately hepatocellular carcinoma. The current approach provides a highly efficient and affordable method to identify causative mutations induced by ENU mutagenesis and animal models relevant to human pathology. |
| Databáze: | OpenAIRE |
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