Acadl-SNP based genotyping assay for long-chain acyl-CoA dehydrogenase deficient mice

The long-chain acyl-CoA dehydrogenase (LCAD) (Acadl=gene; LCAD=protein) deficient mouse model has been important in evaluating the role of mitochondrial fatty acid oxidation of long-chain fatty acids in metabolic disorders. The insertion vector-based gene targeting strategy used to generate this mod...

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Bibliographic Details
Published in:	Molecular genetics and metabolism Vol. 106; no. 1; pp. 62 - 67
Main Authors:	Luther, Rita J., Almodovar, Alvin J.O., Fullerton, Russell, Wood, Philip A.
Format:	Journal Article
Language:	English
Published:	United States Elsevier Inc 01-05-2012
Subjects:	Acadl alleles Acyl-CoA dehydrogenase Acyl-CoA Dehydrogenase, Long-Chain - deficiency Acyl-CoA Dehydrogenase, Long-Chain - genetics Alleles Animal models Animals Base Sequence Disease Models, Animal Fatty acids Fatty Acids - genetics Fatty Acids - metabolism Gene targeting genomics Genotype Genotypes Genotyping Heterozygote Homozygote Humans Insertion Long-chain acyl-CoA dehydrogenase Metabolic disorders Mice Mice, Inbred C57BL Mitochondria Mitochondria - genetics Mitochondria - metabolism Molecular genotyping Molecular Sequence Data Mouse model Oxidation Polymorphism, Single Nucleotide Proteins Single-nucleotide polymorphism SNP genotyping assay Acadl alleles Molecular genotyping Acadl LCAD Long-chain acyl-CoA dehydrogenase Mouse model SNP genotyping assay
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Summary:	The long-chain acyl-CoA dehydrogenase (LCAD) (Acadl=gene; LCAD=protein) deficient mouse model has been important in evaluating the role of mitochondrial fatty acid oxidation of long-chain fatty acids in metabolic disorders. The insertion vector-based gene targeting strategy used to generate this model has made it difficult to distinguish homozygous and heterozygous genotypes containing targeted Acadl alleles in LCAD-deficient mice. Herein, we describe the design and validation of Acadl SNP genotyping methods capable of distinguishing between heterozygous and homozygous LCAD-deficient mice. The Acadl SNP genotyping assays are effective at allelic discrimination of both C57BL/6 and 129 mouse strain-based Acadl alleles under conditions including, both low purity and quantity genomic DNA templates. This makes the method practical and provides the necessary tools for genotyping the LCAD-deficient mouse model. ► Genotyping of Acadl targeted alleles in LCAD-deficient mice has been impractical. ► We have developed Acadl SNP genotyping assays for LCAD-deficient mice. ► The Acadl SNP genotyping assays are effective at both low DNA purity and quantity.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	1096-7192 1096-7206
DOI:	10.1016/j.ymgme.2012.02.009