The services of the Genetics Core will be utilized by both Project 1 and 2. The Genetics Core has several responsibilities for this proposal. The first is the extraction and storage of DNA from whole blood received from the five study sites. The second is the coordination of the genome wide association study (GWAS) and the validation studies. Both the DNA samples and the GWAS information will be used in both Project 1 and Project 2. DNA will also be used for follow-up SNP genotyping within the validation cohort as determined by the results of the GWAS analysis. Additional genotyping of candidate SNPs will be done for Aim 4 of Project 2. Additionally, the Genetics Core will be involved in the isolation of RNA from peripheral white blood cells (WBCs) and conduct gene expression analysis for Project 2. Whole blood will be received from five clinical sites. The Genetics Core will be responsible for extracting DNA from these samples, quality control assessment of DNA samples, sample tracking and sample storage. All samples will have specific sample identification numbers that prevent identification ofthe sample source. As a safety measure, an aliquot of all samples will be stored offsite in Dr. Israni's laboratory (Project 1). DNA samples will be separated into two cohorts, one cohort used for the initial GWAS analysis and the second cohort for SNP validation. These cohorts will contain DNA that has been isolated in the current proposal and samples obtained through this proposal. The majority of the DNA samples isolated through this proposal will be used for the creation of the validation cohort. The Genetics Core will then set up samples for and coordinate the GWAS study which will provide genetic data for Project 1 and 2. The Genetics Core will also set up samples for and coordinate the validation studies for Project 1 and 2. Additional techniques that done by the Genetics Core include candidate SNP analysis for Project 2 and quantitative PCR of expression of candidate genes, also for Project 2. Genotyping for the validation cohort will be conducted in Dr. Israni's laboratory and facilitated by the Genetics Core.
The relevance of the Genetics Core is to facilitate the production of genotypes from patient DNA for the GWAS which will be used by both Project 1 and Project 2. This includes extraction of DNA from patient blood samples, tracking and storing DNA, and coordinating the genotyping. The Genetics Core will also provide additional genotyping of candidate genes and expression analysis for Project 2.
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|Oetting, W S; Schladt, D P; Guan, W et al. (2016) Genomewide Association Study of Tacrolimus Concentrations in African American Kidney Transplant Recipients Identifies Multiple CYP3A5 Alleles. Am J Transplant 16:574-82|
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|Ong, Song; Mannon, Roslyn B (2015) Genomic and proteomic fingerprints of acute rejection in peripheral blood and urine. Transplant Rev (Orlando) 29:60-7|
|Ma, Jun; Divers, Jasmin; Palmer, Nicholette D et al. (2015) Deceased donor multidrug resistance protein 1 and caveolin 1 gene variants may influence allograft survival in kidney transplantation. Kidney Int 88:584-92|
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|Freedman, B I; Julian, B A; Pastan, S O et al. (2015) Apolipoprotein L1 gene variants in deceased organ donors are associated with renal allograft failure. Am J Transplant 15:1615-22|
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