The purpose of the Real-time Quantitative PCR Core is to assist investigators in evaluating gene transfer efficiency and transgene expression in target cells for projects in the Gene Therapy for Metabolic Disorders program. As initially demonstrated by a gene therapy clinical trial (""""""""Lymphocyte gene therapy for Hunter syndrome), this technique fills a very significant scientific need, to accurately quantitate the presence of a gene, and to measure its expression by reverse transcriptase-PCR (RT-PCR) with a high level of sensitivity and reproducability. This core will assist investigators in designing assay systems (paired oligonucleotide primers and an internal TaqMan probe, Black Hole Quencher probe, or SYBR Green), as well as accomplish the actual assays of research specimens. The availability of a core facility accomplishing assays for common biologic systems (e.g., gene and gene products for OTC, MPS I, and MPS VII mouse models) eliminates redundancy and provides a greater level of quality assurance. The new, quantitative technology also provides sensitive methods for """"""""safety assays"""""""" (e.g., for lentiviral vector RCR) that are much faster, reliable and inexpensive in comparison to existing culture approaches. The technique also provides a method of quantitating gene products (RT-PCR) for which there are no existing assays, or for DNA sequences that do not yield a translational or transcriptional end product (as might be needed for transposon vectors).

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
5P01HD032652-10
Application #
7062722
Study Section
Pediatrics Subcommittee (CHHD)
Project Start
Project End
Budget Start
2005-01-01
Budget End
2005-12-31
Support Year
10
Fiscal Year
2005
Total Cost
$66,547
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Type
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Ou, Li; Przybilla, Michael J; Whitley, Chester B (2018) Metabolomics profiling reveals profound metabolic impairments in mice and patients with Sandhoff disease. Mol Genet Metab :
Ou, L; Przybilla, M J; Whitley, C B (2018) SAAMP 2.0: An algorithm to predict genotype-phenotype correlation of lysosomal storage diseases. Clin Genet 93:1008-1014
Ou, Li; Przybilla, Michael J; Whitley, Chester B (2017) Phenotype prediction for mucopolysaccharidosis type I by in silico analysis. Orphanet J Rare Dis 12:125
Hyland, Kendra A; Aronovich, Elena L; Olson, Erik R et al. (2017) Transgene Expression in Dogs After Liver-Directed Hydrodynamic Delivery of Sleeping Beauty Transposons Using Balloon Catheters. Hum Gene Ther 28:541-550
Aronovich, Elena L; Hyland, Kendra A; Hall, Bryan C et al. (2017) Prolonged Expression of Secreted Enzymes in Dogs After Liver-Directed Delivery of Sleeping Beauty Transposons: Implications for Non-Viral Gene Therapy of Systemic Disease. Hum Gene Ther 28:551-564
Ou, Li; Przybilla, Michael J; Whitley, Chester B (2017) Proteomic analysis of mucopolysaccharidosis I mouse brain with two-dimensional polyacrylamide gel electrophoresis. Mol Genet Metab 120:101-110
Verhaart, Ingrid E C; Robertson, Agata; Wilson, Ian J et al. (2017) Prevalence, incidence and carrier frequency of 5q-linked spinal muscular atrophy - a literature review. Orphanet J Rare Dis 12:124
Ou, Li; Przybilla, Michael J; Koniar, Brenda L et al. (2016) Elements of lentiviral vector design toward gene therapy for treating mucopolysaccharidosis I. Mol Genet Metab Rep 8:87-93
Aronovich, Elena L; Hackett, Perry B (2015) Lysosomal storage disease: gene therapy on both sides of the blood-brain barrier. Mol Genet Metab 114:83-93
Satzer, David; DiBartolomeo, Christina; Ritchie, Michael M et al. (2015) Assessment of dysmyelination with RAFFn MRI: application to murine MPS I. PLoS One 10:e0116788

Showing the most recent 10 out of 92 publications