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CPT2Carnitine Palmitoyl Transferase 2
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Lipid metabolism: The following genes were categorized into the lipid metabolism pathway: PLTP (lipid transport); SCD (lipogenesis); liver X receptor a, nuclear receptor subfamily 1, group H, member (LR1H3) (cholesterol metabolism); FABP3 (fatty acid transport); CPT2, ACADL, and ACOX2 (fatty acid oxidation); and FABP4 (adipocyte differentiation).
We also found that the expression levels of the CPT2, ACADL, and ACOX2 genes were increased in the high-marbled group in the present study.
Activation of PPARG promotes terminal differentiation through the induction of a range of genes important for TG formation through fatty acid oxidation among downstream pathways involving CPT2, ACADL, and ACOX2.
Screening for mutations in CPT2 has been performed using DNA isolated from biopsied muscle tissue (9), venous blood (10), lymphoblasts (11), or fibroblasts (8).
Although the methods for DNA isolation, PCR amplification, and ASO analysis developed previously by our laboratory are effective for detecting known mutations in the CPT2 gene (9), they were too costly and laborious for application to large family studies or population screening.
Twenty additional genomic DNA specimens were prepared from wet tissue (whole blood, muscle, lymphoblasts, or fibroblasts), previously screened for CPT2 mutations (9) and used as controls (50 mg/L) for the PCR and ASO optimization experiments.
Subsequent analysis for additional mutations in the CPT2 gene indicated that this degree of sensitivity is similar for all of the ASOs used with the same molar ratio of biotinylated oligonucleotides to amplified targets (data not shown).
We have developed a protocol that optimizes and shortens the CPT2 mutation screening procedure from specimen collection to mutation detection, producing a specific and sensitive assay using DNA from dried blood spots.