The Molecular Core of the YCCMD provides support for a number of specialized methodologies and technical assistance in essential molecular techniques. The creation of animal models of gene dysregulation and the characterization of target gene expression is fully supported the Molecular Core, with further analysis both in vivo and in vitro provided by the Cell and Physiology Cores.
Aim 1. To provide as services, a) the construction of transgenes and gene disruption vectors, and b) the identification of genetically altered or naturally occurring mutant mice. This includes constitutive transgenes, inducible transgenes using the tetracycline transactivator system, conventional gene ablation vectors, conditional gene targeting vectors (using Cre-loxP or Flp-frf approaches), gene knock-in with expression tags (IRES-LacZ or -GFP) and inducible gene targeting (with Cre-ER or Cre-PR fusions). Genotyping is carried out by PCR and/or Southern blot analysis of tail DMA and/or ES cell DMA.
Aim 2. To provide as services, a) the localization of gene expression by in situ hybridization, and b) the quantitation of gene expression by quantitative, real-time RT-PCR. For expression localization, this includes the dissection and fixation of tissues, embedding in paraffin or OCT, sectioning by microtome or cryostat, preparation of riboprobes or oligoprobes, hybridization, emulsion autoradiography and image analysis. For quantitation of gene expression, this includes tissue dissection, RNA preparation, RT primer selection and PCR primer design.
Aim 3. To provide technical assistance, training and resources for essential molecular methods, including: RNA and genomic DNA preparation and analysis;stable or transient cell transfection; reporter gene analysis;PCR-related techniques, such as mutagenesis, cloning, overlap extension and inverse PCR;siRNA design, Dicer pools and shRNA (short hairpin) vectors and viruses for projects that require the suppression of gene expression by RNA interference;and BAG recombineering.

Public Health Relevance

The mission of the Molecular Core is to support the creation and analysis of animal models relevant to musculoskeletal disease and to facilitate the introduction and use of a range of molecular genetic methods by member investigators.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Center Core Grants (P30)
Project #
5P30AR046032-15
Application #
8452190
Study Section
Special Emphasis Panel (ZAR1-CHW-G)
Project Start
Project End
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
15
Fiscal Year
2013
Total Cost
$189,286
Indirect Cost
$76,285
Name
Yale University
Department
Type
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
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Fretz, Jackie A; Nelson, Tracy; Velazquez, Heino et al. (2014) Early B-cell factor 1 is an essential transcription factor for postnatal glomerular maturation. Kidney Int 85:1091-102
McCarthy, Thomas L; Yun, Zhong; Madri, Joseph A et al. (2014) Stratified control of IGF-I expression by hypoxia and stress hormones in osteoblasts. Gene 539:141-51
Scheller, Erica L; Troiano, Nancy; Vanhoutan, Joshua N et al. (2014) Use of osmium tetroxide staining with microcomputerized tomography to visualize and quantify bone marrow adipose tissue in vivo. Methods Enzymol 537:123-39

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